Compounds useful in hiv therapy

ABSTRACT

The invention relates to a compound having the structure: 
     
       
         
         
             
             
         
       
         
         
           
             or a pharmaceutically acceptable salt thereof, along with pharmaceutical compositions and therapeutic methods thereof.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation application of Ser. No. 16/865,678filed May 4, 2020, which is an application filed under 35 USC 111(a)claiming priority to PCT Application No. PCT/IB2020/051878 filed Mar. 4,2020, which claims priority to U.S. Provisional Ser. No. 62/814,316filed Mar. 6, 2019, the disclosure of which is incorporated by referenceherein in its entirety.

FIELD OF THE INVENTION

The present invention relates to compounds, pharmaceutical compositions,and methods of use thereof in connection with individuals infected withHIV. In particular, such methods of use encompass e.g., methods fortreating HIV and methods of preventing HIV.

BACKGROUND OF THE INVENTION

Human immunodeficiency virus type 1 (HIV-1) infection leads to thecontraction of acquired immune deficiency disease (AIDS). The number ofcases of HIV continues to rise, and currently an estimated overthirty-five million individuals worldwide suffer from HIV infectione.g.,http://www.sciencedirect.com/science/article/pii/S235230181630087X?via%3Dihub

Presently, long-term suppression of viral replication withantiretroviral drugs is the only option for treating HIV-1 infection.Indeed, the U.S. Food and Drug Administration has approved twenty-fivedrugs over six different inhibitor classes, which have been shown togreatly increase patient survival and quality of life. However,additional therapies are still believed to be required due to a numberof issues including, but not limited to undesirable drug-druginteractions; drug-food interactions; non-adherence to therapy; drugresistance due to mutation of the enzyme target; and inflammationrelated to the immunologic damage caused by the HIV infection.

Currently, almost all HIV positive patients are treated with therapeuticregimens of antiretroviral drug combinations termed, highly activeantiretroviral therapy (“HAART”). However, HAART therapies are oftencomplex because a combination of different drugs must be administeredoften daily to the patient to avoid the rapid emergence ofdrug-resistant HIV-1 variants. Despite the positive impact of HAART onpatient survival, drug resistance can still occur and the survival andquality of life are not normalized as compared to uninfected persons[Lohse Ann Intern Med 2007 146; 87-95]. Indeed, the incidence of severalnon-AIDS morbidities and mortalities, such as cardiovascular disease,frailty, and neurocognitive impairment, are increased inHAART-suppressed, HIV-infected subjects [Deeks Annu Rev Med 2011;62:141-155]. This increased incidence of non-AIDS morbidity/mortalityoccurs in the context of, and is potentially caused by, elevatedsystemic inflammation related to the immunologic damage caused by HIVinfection [Hunt J Infect Dis 2014][Byakagwa J Infect Dis 2014][Tenorio JInfect Dis 2014].

Modern antiretroviral therapy (ART) has the ability to effectivelysuppress HIV replication and improve health outcomes for HIV-infectedpersons, but is believed to not be capable of completely eliminating HIVviral reservoirs within the individual. HIV genomes can remain latentwithin mostly immune cells in the infected individual and may reactivateat any time, such that after interruption of ART, virus replicationtypically resumes within weeks. In a handful of individuals, the size ofthis viral reservoir has been significantly reduced and upon cessationof ART, the rebound of viral replication has been delayed [Henrich T J JInfect Dis 2013][Henrich T J Ann Intern Med 2014]. In one case, theviral reservoir was eliminated during treatment of leukemia and no viralrebound was observed during several years of follow-up [Hutter G N EnglJ Med 2009]. These examples suggest the concept that reduction orelimination of the viral reservoir may be possible and can lead to viralremission or cure. As such, ways have been pursued to eliminate theviral reservoir, by direct molecular means, including excision of viralgenomes with CRISPR/Cas9 systems, or to induce reactivation of thelatent reservoir during ART so that the latent cells are eliminated.Induction of the latent reservoir typically results in either directdeath of the latently infected cell or killing of the induced cell bythe immune system after the virus is made visible. As this is performedduring ART, viral genomes produced are believed to not result in theinfection of new cells and the size of the reservoir may decay.

HAART therapies are often complex because a combination of differentdrugs must be administered often daily to the patient to avoid the rapidemergence of drug-resistant HIV-1 variants. Despite the positive impactof HAART on patient survival, drug resistance can still occur.

Current guidelines recommend that therapy includes three fully activedrugs. See e.g. https://aidsinfo.nih.gov/quidelines. Additionally, twodrug combinations may be employed as therapeutic regimens. Typically,first-line therapies combine two to three drugs targeting the viralenzymes reverse transcriptase and integrase. It is believed thatsustained successful treatment of HIV-1-infected patients withantiretroviral drugs employ the continued development of new andimproved drugs that are effective against HIV strains that have formedresistance to approved drugs. For example an individual on a regimencontaining 3TC/FTC (lamivudine/emtricitabine) may select for the M184Vmutation that reduces susceptibility to these drugs by >100 fold. See eg., https://hivdb.stanford.edu/dr-summary/resistance-notes/NRTI

Another way to potentially address preventing formation of mutations isto increase patient adherence to a drug regimen. One manner that may beemployed to accomplish this is by reducing the dosing frequency. Forparenteral administration, it is believed to be advantageous to havedrug substances with high lipophilicity in order to reduce solubilityand limit the release rate within interstitial fluid. However, mostnucleoside reverse transcriptase inhibitors are hydrophilic therebypotentially limiting their use as long acting parenteral agents.

There remains a need for compounds which may address the shortcomingsset forth above.

SUMMARY OF THE INVENTION

In one aspect, the invention provides a compound of the formula (I):

wherein:

R¹ is:

wherein:

X is selected from the group consisting of NH₂, F and Cl;

R⁵ is selected from the group consisting of H and (C₁-C₁₄) alkyl;

R⁶ is selected from the group consisting of H and —(C═O)—(C₁-C₁₄) alkyl;

R² is selected from the group consisting of (C₁-C₂₄) alkyl;(CH₂)_(n1)—O—(CH₂CH₂O)_(n2)—(C₁-C₁₄ alkyl) where n1 and n2 are integersindependently selected from 1-4; —R⁷—NH—(C═O)—R⁸ wherein R⁷ may be(C₁-C₁₄) alkyl and R^(a) may be independently selected from H and(C₁-C₁₄) alkyl; —R⁹—(C₆-C₁₄) aryl, wherein R⁹ is a bond or (C₁-C₆)alkyl; —R¹⁰—(C₃-C₁₄) cycloalkyl, wherein R¹⁰ is a bond or (C₁-C₆) alkyl;—(C₁-C₂₀) alkylene-(C═O)—O—R¹¹ wherein R¹¹ may be selected from H and(C₁-C₂₀)alkyl;

and;

R³ is selected from the group consisting of H, —(C═O)—(C₁-C₂₄) alkyl;—(C═O)—O—(C₁-C₂₄) alkyl; and C₃-C₁₄ cycloalkyl; or

R² and R³ join together to form a C₃ to C₂₈ cyclic structure; and withthe proviso that when R² is (C₁-C₁₄ alkyl) at least one of R³, R⁵ and R⁶is not H.

In another aspect, the invention provides pharmaceutical compositionscomprising a compound of Formula (I) or a pharmaceutically acceptablesalt thereof and an excipient.

In another aspect, the invention provides a method of treating an HIVinfection in a subject at risk for developing an HIV infection,comprising administering to the subject a compound of Formula (I), or apharmaceutically acceptable salt thereof.

In another aspect, the invention provides a method of preventing an HIVinfection in a subject at risk for developing an HIV infection,comprising administering to the subject a compound of Formula (I), or apharmaceutically acceptable salt thereof.

In another aspect, there is provided a compound of Formula (I) or apharmaceutically acceptable salt thereof for use in therapy.

In another aspect, there is provided a compound of Formula (I) or apharmaceutically acceptable salt thereof for use in treating an HIVinfection.

In another aspect, there is provided a compound of Formula (I) or apharmaceutically acceptable salt thereof for use in preventing an HIVinfection.

In another aspect, there is provided the use of a compound of Formula(I) or a pharmaceutically acceptable salt thereof in the manufacture ofa medicament for treating an HIV infection.

In another aspect, there is provided the use of a compound of Formula(I) or a pharmaceutically acceptable salt thereof in the manufacture ofa medicament for preventing an HIV infection.

These and other aspects are encompassed by the invention as set forthherein.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A illustrates an IC₅₀ curve shift from t=0 to t=48 h for EFdA.

FIG. 1B illustrates an IC₅₀ curve shift from t=0 to t=48 h for Example18 of the present invention.

FIG. 2 illustrates the mean plasma concentration-time profiles of EFdAafter subcutaneous dosing at 20 mg/kg in Wistar Han rats (N=3/timepoint)

FIG. 3 illustrates the mean plasma concentration-time profiles of EFdAafter intramuscular dosing at 20 mg/kg in Wistar Han rats (N=3/timepoint)

FIG. 4 illustrates the mean plasma concentration-time profiles ofExample 11 and EFdA after single IM injection of Example 11 at 20 mg/kgequivalent of EFdA in male Wistar Han rats (N=3/time point)

FIG. 5 illustrates the mean plasma concentration-time profiles ofExample 17 and EFdA after single IM injection of Example 17 at 20 mg/kgequivalent of EFdA in male Wistar Han rats (N=3/time point)

FIG. 6 illustrates the mean plasma concentration-time profiles ofExample 18, EFdA and Met1 after single IM injection of Example 18 at 20mg/kg equivalent of EFdA in male Wistar Han rats (N=3/time point)

FIG. 7 illustrates the mean plasma concentration-time profiles ofExample 31, EFdA and Met1 after single IM injection of Example 31 at 20mg/kg equivalent of EFdA in male Wistar Han rats (N=3/time point)

FIG. 8 illustrates the mean plasma concentration-time profiles ofExample 32, EFdA, Met1 and Met2 after single IM injection of Example 32at 20 mg/kg equivalent of EFdA in male Wistar Han rats (N=3/time point)

FIG. 9 illustrates the mean plasma concentration-time profiles ofExample 37, EFdA, Met1 and Met2 after single IM injection of Example 37at 20 mg/kg equivalent of EFdA in male Wistar Han rats (N=3/time point)

FIG. 10 illustrates the mean plasma concentration-time profiles ofExample 38, EFdA, Met1 and Met2 after single IM injection of Example 38at 20 mg/kg equivalent of EFdA in male Wistar Han rats (N=3/time point)

FIG. 11 illustrates the mean plasma concentration-time profiles ofExample 39, EFdA, Met1 and Met2 after single IM injection of Met2 at 20mg/kg equivalent of EFdA in male Wistar Han rats (N=3/time point)

FIG. 12 illustrates the mean plasma concentration-time profiles ofExample 40 and EFdA after single IM injection of Example 40 at 20 mg/kgequivalent of EFdA in male Wistar Han rats (N=3/time point)

FIG. 13 illustrates the mean plasma concentration-time profiles ofExample 42, EFdA, Met1 and Met2 after single IM injection of Example 42at 20 mg/kg equivalent of EFdA in male Wistar Han rats (N=3/time point)

FIG. 14 illustrates the mean plasma concentration-time profiles ofExample 45, EFdA, Met1 and Met2 after single IM injection of Example 45at 20 mg/kg equivalent of EFdA in male Wistar Han rats (N=3/time point)

FIG. 15 illustrates the mean plasma concentration-time profiles ofExample 48 and EFdA after single IM injection of Example 48 at 20 mg/kgequivalent of EFdA in male Wistar Han rats (N=3/time point)

FIG. 16 illustrates the mean plasma concentration-time profiles ofExample 50 and EFdA after single IM injection of Example 50 at 20 mg/kgequivalent of EFdA in male Wistar Han rats (N=3/time point)

FIG. 17 illustrates the mean plasma concentration-time profiles ofExample 58 and EFdA after single IM injection of Example 58 at 20 mg/kgequivalent of EFdA in male Wistar Han rats (N=3/time point)

FIG. 18 illustrates the mean plasma concentration-time profiles ofExample 59 and EFdA after single IM injection of Example 59 at 20 mg/kgequivalent of EFdA in male Wistar Han rats (N=3/time point)

FIG. 19 illustrates the mean plasma concentration-time profiles ofExample 61 and EFdA after single IM injection of Example 61 at 20 mg/kgequivalent of EFdA in male Wistar Han rats (N=3/time point)

FIG. 20 illustrates the mean plasma concentration-time profiles ofExample 62 and EFdA after single IM injection of Example 62 at 20 mg/kgin male Wistar Han rats (N=3/time point)

FIG. 21 illustrates the mean plasma concentration-time profiles ofExample 63 and EFdA after single IM injection of Example 63 at 20 mg/kgequivalent of EFdA in male Wistar Han rats (N=3/time point)

FIG. 22 illustrates the mean plasma concentration-time profiles ofExample 64 and EFdA after single IM injection of Example 64 at 20 mg/kgequivalent of EFdA in male Wistar Han rats (N=3/time point)

FIG. 23 illustrates the mean plasma concentration-time profiles ofExample 65 and EFdA after single IM injection of Example 65 at 20 mg/kgequivalent of EFdA in male Wistar Han rats (N=3/time point)

FIG. 24 illustrates the mean plasma concentration-time profiles ofexample 66 and EFdA after single IM injection of Example 66 at 20 mg/kgequivalent of EFdA in male Wistar Han rats (N=3/time point)

FIG. 25 illustrates the mean plasma concentration-time profiles ofExample 18 and EFdA after subcutaneous dosing Example 18 at 20 mgequivalent of EFdA/kg in Wistar Han rats (N=3/time point)

FIG. 26 illustrates the mean plasma concentration-time profiles ofExample 18 and EFdA after intramuscular dosing Example 18 at 20 mgequivalent of EFdA/kg in Wistar Han rats (N=3/time point)

FIG. 27 illustrates the mean plasma concentration-time profiles ofExample 18 and EFdA after subcutaneous dosing Example 18 at 5 mgequivalent of EFdA/kg in Beagle Dogs (N=3/time point)

FIG. 28 illustrates the mean plasma concentration-time profiles ofExample 18 and EFdA after intramuscular dosing Example 18 at 5 mgequivalent of EFdA/kg in Beagle Dogs (N=3/time point)

FIG. 29 illustrates the mean plasma concentration-time profiles ofExample 18 and EFdA after subcutaneous dosing Example 18 at 5 mgequivalent of EFdA/kg in Cynomolgus Monkey (N=3/time point)

FIG. 30 illustrates the mean plasma concentration-time profiles ofExample 18 and EFdA after intramuscular dosing Example 18 at 5 mgequivalent of EFdA/kg in Cynomolgus Monkey (N=3/time point)

DETAILED DESCRIPTION OF REPRESENTATIVE EMBODIMENTS

Throughout this application, references are made to various embodimentsrelating to compounds, compositions, and methods. The variousembodiments described are meant to provide a variety of illustrativeexamples and should not be construed as descriptions of alternativespecies. Rather it should be noted that the descriptions of variousembodiments provided herein may be of overlapping scope. The embodimentsdiscussed herein are merely illustrative and are not meant to limit thescope of the present invention.

It is to be understood that the terminology used herein is for thepurpose of describing particular embodiments only and is not intended tolimit the scope of the present invention. In this specification and inthe claims that follow, reference will be made to a number of terms thatshall be defined to have the following meanings, unless otherwise noted.

As used herein unless otherwise specified and subject to otherembodiments set forth herein, “alkyl” e.g., refers to a monovalentsaturated aliphatic hydrocarbyl group having e.g., from 1 to 24 carbonatoms, from 1 to 20 carbon atoms, from 1 to 14 carbon atoms or, from 1to 6 carbon atoms. “(C_(x)-C_(y))alkyl” refers to alkyl groups havingfrom x to y carbon atoms. The term “alkyl” includes, by way of example,linear and branched hydrocarbyl groups such as, e.g., methyl (CH₃—),ethyl (CH₃CH₂—), n-propyl (CH₃CH₂CH₂—), isopropyl ((CH₃)₂CH—), n-butyl(CH₃CH₂CH₂CH₂—), isobutyl ((CH₃)₂CHCH₂—), sec-butyl ((CH₃)(CH₃CH₂)CH—),t-butyl ((CH₃)₃C—), n-pentyl (CH₃CH₂CH₂CH₂CH₂—), and neopentyl((CH₃)₃CCH₂—). For the purposes of the invention, alkyl may beinterpreted to encompass alkylene groups defined herein.

Subject to various embodiments set forth herein, “Alkylene” or“alkylene” refers to divalent e.g., saturated aliphatic hydrocarbylgroups having from 1 to 6 carbon atoms. The alkylene groups includebranched and straight chain hydrocarbyl groups. For example,“(C₁-C₆)alkylene” is meant to include methylene, ethylene, propylene,2-methypropylene, dimethylethylene, pentylene, and so forth. As such,the term “propylene” could be exemplified by the following structure:

Likewise, the term “dimethylbutylene” could be exemplified by any of thefollowing three structures or more:

p, or

Furthermore, the term “(C₁-C₆)alkylene” is meant to include suchbranched chain hydrocarbyl groups as cyclopropylmethylene, which couldbe exemplified by the following structure:

“Alkenyl” refers to a linear or branched hydrocarbyl group having, e.g.,from 2 to 10 carbon atoms and in some embodiments from 2 to 6 carbonatoms or 2 to 4 carbon atoms and having at least 1 site of vinylunsaturation (>C═C<). For example, (C_(x)-C_(y))alkenyl refers toalkenyl groups having from x to y carbon atoms and is meant to includefor example, ethenyl, propenyl, isopropylene, 1,3-butadienyl, and thelike.

“Alkynyl” refers to a linear monovalent hydrocarbon radical or abranched monovalent hydrocarbon radical containing at least one triplebond. The term “alkynyl” is also meant to include those hydrocarbylgroups having one triple bond and one double bond. For example,(C₂-C₆)alkynyl is meant to include ethynyl, propynyl, and the like.

“Alkoxy” refers to the group —O-alkyl wherein alkyl is defined herein,e.g., C₁ to C₆ alkoxy. Alkoxy includes, by way of example, methoxy,ethoxy, n-propoxy, isopropoxy, n-butoxy, t-butoxy, sec-butoxy, andn-pentoxy.

“Aryl” refers to an aromatic group of from e.g., 6 to 14 carbon atoms orpreferably 5 to 6 carbon atoms and no ring heteroatoms and having asingle ring (e.g., phenyl) or multiple condensed (fused) rings (e.g.,naphthyl or anthryl). For multiple ring systems, including fused,bridged, and spiro ring systems having aromatic and non-aromatic ringsthat have no ring heteroatoms, the term “Aryl” or “Ar” applies when thepoint of attachment is at an aromatic carbon atom (e.g., 5,6,7,8tetrahydronaphthalene-2-yl is an aryl group as its point of attachmentis at the 2-position of the aromatic phenyl ring). As set forth herein,aryl groups may be substituted.

“AUC” refers to the area under the plot of plasma concentration of drug(not logarithm of the concentration) against time after drugadministration.

“Cycloalkyl” refers to a saturated or partially saturated cyclic groupof from e.g, 3 to 14 carbon atoms and no ring heteroatoms and having asingle ring or multiple rings including fused, bridged, and spiro ringsystems. For multiple ring systems having aromatic and non-aromaticrings that have no ring heteroatoms, the term “cycloalkyl” applies whenthe point of attachment is at a non-aromatic carbon atom (e.g.5,6,7,8,-tetrahydronaphthalene-5-yl). The term “cycloalkyl” includescycloalkenyl groups, such as cyclohexenyl. Examples of cycloalkyl groupsinclude, for instance, adamantyl, cyclopropyl, cyclobutyl, cyclohexyl,cyclopentyl, cyclooctyl, cyclopentenyl, and cyclohexenyl. Examples ofcycloalkyl groups that include multiple bicycloalkyl ring systems arebicyclohexyl, bicyclopentyl, bicyclooctyl, and the like. Two suchbicycloalkyl multiple ring structures are exemplified and named below:

“(C_(u)-C_(v))cycloalkyl” refers to cycloalkyl groups having u to vcarbon atoms.

“EC₅₀” refers to the concentration of a drug that gives half-maximalresponse.

“IC₅₀” refers to the half-maximal inhibitory concentration of a drug.Sometimes, it is also converted to the pIC₅₀ scale (−log IC₅₀), in whichhigher values indicate exponentially greater potency.

“Haloalkyl” refers to substitution of an alkyl group with 1 to 3 halogroups (e.g., bifluoromethyl or trifluoromethyl).

“Heteroaryl” refers to an aromatic group of from 1 to 14 carbon atoms,e.g., 6 heteroatoms selected from oxygen, nitrogen, and sulfur andincludes single ring (e.g. imidazolyl) and (e.g. benzimidazol-2-yl andbenzimidazol-6-yl). For multiple ring systems, including fused, bridged,and spiro ring systems having aromatic and non-aromatic rings, the term“heteroaryl” applies if there is at least one ring heteroatom and thepoint of attachment is at an atom of an aromatic ring (e.g.1,2,3,4-tetrahydroquinolin-6-yl and 5,6,7,8-tetrahydroquinolin-3-yl). Insome embodiments, the nitrogen and/or the sulfur ring atom(s) of theheteroaryl group are optionally oxidized to provide for the N-oxide(N→O), sulfinyl, or sulfonyl moieties. More specifically the termheteroaryl includes, but is not limited to, pyridyl, furanyl, thienyl,thiazolyl, isothiazolyl, triazolyl, imidazolyl, imidazolinyl,isoxazolyl, pyrrolyl, pyrazolyl, pyridazinyl, pyrimidinyl, purinyl,phthalazyl, naphthylpryidyl, benzofuranyl, tetrahydrobenzofuranyl,isobenzofuranyl, benzothiazolyl, benzoisothiazolyl, benzotriazolyl,indolyl, isoindolyl, indolizinyl, dihydroindolyl, indazolyl, indolinyl,benzoxazolyl, quinolyl, isoquinolyl, quinolizyl, quianazolyl,quinoxalyl, tetrahydroquinolinyl, isoquinolyl, quinazolinonyl,benzimidazolyl, benzisoxazolyl, benzothienyl, benzopyridazinyl,pteridinyl, carbazolyl, carbolinyl, phenanthridinyl, acridinyl,phenanthrolinyl, phenazinyl, phenoxazinyl, phenothiazinyl, andphthalimidyl.

“Heterocycle” refers “Heterocyclic” or “heterocycle” or“heterocycloalkyl” or “heterocyclyl” refers to a saturated or partiallysaturated cyclic group having from 1 to 14 carbon atoms and from 1 to 6heteroatoms selected from nitrogen, sulfur, phosphorus or oxygen andincludes single ring and multiple ring systems including fused, bridged,and spiro ring systems. For multiple ring systems having aromatic and/ornon-aromatic rings, the terms “heterocyclic”, “heterocycle”,“heterocycloalkyl”, or “heterocyclyl” apply when there is at least onering heteroatom and the point of attachment is at an atom of anon-aromatic ring (e.g. 1,2,3,4-tetrahydroquinoline-3-yl,5,6,7,8-tetrahydroquinoline-6-yl, and decahydroquinolin-6-yl). In oneembodiment, the nitrogen, phosphorus and/or sulfur atom(s) of theheterocyclic group are optionally oxidized to provide for the N-oxide,phosphinane oxide, sulfinyl, sulfonyl moieties. More specifically theheterocyclyl includes, but is not limited to, tetrahydropyranyl,piperidinyl, piperazinyl, 3-pyrrolidinyl, 2-pyrrolidon-1-yl,morpholinyl, and pyrrolidinyl. A prefix indicating the number of carbonatoms (e.g., C₃-C₁₀) refers to the total number of carbon atoms in theportion of the heterocyclyl group exclusive of the number ofheteroatoms.

“Fused heterocyclic” or “fused heterocycle” refer to a 3 to 10 membercyclic substituent formed by the replacement of two hydrogen atoms atdifferent carbon atoms in a cycloalkyl ring structure, as exemplified bythe following structure wherein the cycloalkyl group shown here containsbonds marked with wavy lines which are bonded to carbon atoms that aresubstituted with a fused heterocyclic group:

“Cyclic structure” refers to a series of carbon atoms connected to forma ring. In various embodiments, the cyclic structure may include e.g.,cycloalkyl and heterocyclic groups. As noted herein, “C_(x) to C_(y)cyclic structure” refers to a cyclic structure wherein x and y are setforth such that the structure may contain x to y carbon atoms. Such astructure may be optionally substituted as set forth herein.

“Compound”, “compounds”, “chemical entity”, and “chemical entities” asused herein refers to a compound encompassed by the generic formulae(I), (Ia), (II) and (IIa) disclosed herein, any subgenus of thesegeneric formulae, and any forms of the compounds within the generic andsubgeneric formulae, including the racemates, stereoisomers, andtautomers of the compound or compounds.

The term “heteroatom” means nitrogen, oxygen, or sulfur and includes anyoxidized form of phosphorus, nitrogen, such as N(O) {N⁺—O⁻} and sulfursuch as S(O) and S(O)₂, and the quaternized form of any basic nitrogen.

“Oxo” refers to a (═O) group.

“Polymorphism” refers to when two or more clearly different phenotypesexist in the same population of a species where the occurrence of morethan one form or morph. In order to be classified as such, morphs mustoccupy the same habitat at the same time and belong to a panmicticpopulation (one with random mating).

“Protein binding” refers to the binding of a drug to proteins in bloodplasma, tissue membranes, red blood cells and other components of blood.

“Protein shift” refers to determining a binding shift by comparing theEC₅₀ values determined in the absence and presence of human serum.

“Racemates” refers to a mixture of enantiomers. In an embodiment of theinvention, the compounds of Formulas I, Ia, II and IIa orpharmaceutically acceptable salts thereof, are enantiomerically enrichedwith one enantiomer wherein all of the chiral carbons referred to are inone configuration. In general, reference to an enantiomerically enrichedcompound or salt, is meant to indicate that the specified enantiomerwill comprise more than 50% by weight of the total weight of allenantiomers of the compound or salt.

“Solvate” or “solvates” of a compound refer to those compounds, asdefined above, which are bound to a stoichiometric or non-stoichiometricamount of a solvent. Solvates of a compound includes solvates of allforms of the compound. In certain embodiments, solvents are volatile,non-toxic, and/or acceptable for administration to humans in traceamounts. Suitable solvates include water.

“Stereoisomer” or “stereoisomers” refer to compounds that differ in thechirality of one or more stereocenters. Stereoisomers includeenantiomers and diastereomers.

“Tautomer” refer to alternate forms of a compound that differ in theposition of a proton, such as enol-keto and imine-enamine tautomers, orthe tautomeric forms of heteroaryl groups containing a ring atomattached to both a ring —NH— moiety and a ring ═N— moiety such aspyrazoles, imidazoles, benzimidazoles, triazoles, and tetrazoles.

The term ‘atropisomer’ refers to a stereoisomer resulting from an axisof asymmetry. This can result from restricted rotation about a singlebond where the rotational barrier is high enough to allowdifferentiation of the isomeric species up to and including completeisolation of stable non-interconverting diastereomer or enantiomericspecies. One skilled in the art will recognize that upon installing anonsymmetrical R^(x) to core, the formation of atropisomers is possible.In addition, once a second chiral center is installed in a givenmolecule containing an atropisomer, the two chiral elements takentogether can create diastereomeric and enantiomeric stereochemicalspecies. Depending upon the substitution about the C_(x) axis,interconversion between the atropisomers may or may not be possible andmay depend on temperature. In some instances, the atropisomers mayinterconvert rapidly at room temperature and not resolve under ambientconditions. Other situations may allow for resolution and isolation butinterconversion can occur over a period of seconds to hours or even daysor months such that optical purity is degraded measurably over time. Yetother species may be completely restricted from interconversion underambient and/or elevated temperatures such that resolution and isolationis possible and yields stable species. When known, the resolvedatropisomers were named using the helical nomenclature. For thisdesignation, only the two ligands of highest priority in front andbehind the axis are considered. When the turn priority from the frontligand 1 to the rear ligand 1 is clockwise, the configuration is P, ifcounterclockwise it is M.

“As used herein, the term “pharmaceutically acceptable” refers to thosecompounds, materials, compositions, and dosage forms which are, withinthe scope of sound medical judgment, suitable for use in contact withthe tissues of human beings and animals without excessive toxicity,irritation, or other problem or complication. “Pharmaceuticallyacceptable salt” refers to pharmaceutically acceptable salts derivedfrom a variety of organic and inorganic counter ions well known in theart and include, by way of example only, sodium, potassium, calcium,magnesium, ammonium, and tetraalkylammonium, and when the moleculecontains a basic functionality, salts of organic or inorganic acids,such as hydrochloride, hydrobromide, tartrate, mesylate, acetate,maleate, and oxalate. Suitable salts include those described in P.Heinrich Stahl, Camille G. Wermuth (Eds.), Handbook of PharmaceuticalSalts Properties, Selection, and Use; 2002. Pharmaceutically acceptablesalts may be prepared in situ during the final isolation andpurification of the compound, or by separately reacting the purifiedcompound in its free acid or free base form with a suitable base oracid, respectively. Further to the above, illustrative pharmaceuticallyacceptable acid salts of the compounds of the present invention can beprepared from the following acids, including, without limitation formic,acetic, propionic, benzoic, succinic, glycolic, gluconic, lactic,maleic, malic, tartaric, citric, nitic, ascorbic, glucuronic, maleic,fumaric, pyruvic, aspartic, glutamic, benzoic, hydrochloric,hydrobromic, hydroiodic, isocitric, trifluoroacetic, pamoic, propionic,anthranilic, mesylic, oxalacetic, oleic, stearic, salicylic,p-hydroxybenzoic, nicotinic, phenylacetic, mandelic, embonic (pamoic),methanesulfonic, phosphoric, phosphonic, ethanesulfonic,benzenesulfonic, pantothenic, toluenesulfonic, 2-hydroxyethanesulfonic,sulfanilic, sulfuric, salicylic, cyclohexylaminosulfonic, algenic,p-hydroxybutyric, galactaric and galacturonic acids. Preferredpharmaceutically acceptable salts include the salts of hydrochloric acidand trifluoroacetic acid. Illustrative pharmaceutically acceptableinorganic base salts of the compounds of the present invention includemetallic ions. More preferred metallic ions include, but are not limitedto, appropriate alkali metal salts, alkaline earth metal salts and otherphysiological acceptable metal ions. Salts derived from inorganic basesinclude aluminum, ammonium, calcium, copper, ferric, ferrous, lithium,magnesium, manganic salts, manganous, potassium, sodium, zinc, and thelike and in their usual valences. Exemplary base salts include aluminum,calcium, lithium, magnesium, potassium, sodium and zinc. Other exemplarybase salts include the ammonium, calcium, magnesium, potassium, andsodium salts. Still other exemplary base salts include, for example,hydroxides, carbonates, hydrides, and alkoxides including NaOH, KOH,Na₂CO₃, K₂CO₃, NaH, and potassium-t-butoxide. Salts derived frompharmaceutically acceptable organic non-toxic bases include salts ofprimary, secondary, and tertiary amines, including in part,trimethylamine, diethylamine, N, N′-dibenzylethylenediamine,chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine(N-methylglucamine) and procaine; substituted amines including naturallyoccurring substituted amines; cyclic amines; quaternary ammoniumcations; and basic ion exchange resins, such as arginine, betaine,caffeine, choline, N,N-dibenzylethylenediamine, diethylamine,2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine,ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine,glucosamine, histidine, hydrabamine, isopropylamine, lysine,methylglucamine, morpholine, piperazine, piperidine, polyamine resins,procaine, purines, theobromine, triethylamine, trimethylamine,tripropylamine, tromethamine and the like. All of the above salts can beprepared by those skilled in the art by conventional means from thecorresponding compound of the present invention. For example, thepharmaceutically acceptable salts of the present invention can besynthesized from the parent compound which contains a basic or acidicmoiety by conventional chemical methods. Generally, such salts can beprepared by reacting the free acid or base forms of these compounds witha stoichiometric amount of the appropriate base or acid in water or inan organic solvent, or in a mixture of the two; generally, nonaqueousmedia like ether, ethyl acetate, ethanol, isopropanol, or acetonitrileare preferred. The salt may precipitate from solution and be collectedby filtration or may be recovered by evaporation of the solvent. Thedegree of ionisation in the salt may vary from completely ionised toalmost non-ionised. Lists of suitable salts are found in Remington'sPharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa.,1985, p. 1418, the disclosure of which is hereby incorporated byreference only with regards to the lists of suitable salts.

“Patient” or “subject” refers to mammals and includes humans andnon-human mammals. Most preferably, a “patient” is construed to refer tohumans.

Treating” or “treatment” of a disease in a patient refers to 1)preventing the disease from occurring in a patient that is predisposedor does not yet display symptoms of the disease; 2) inhibiting thedisease or arresting its development; or 3) ameliorating or causingregression of the disease.

Prevention or “preventing” a disease in a patient refers to theprophylactic administration of a drug to substantially diminish thelikelihood or severity of a disorder or biological manifestationthereof, or to delay the onset of such disorder or biologicalmanifestation thereof.

Where specific compounds or generic formulas are drawn that havearomatic rings, such as aryl or heteroaryl rings, then it will beunderstood by one of still in the art that the particular aromaticlocation of any double bonds are a blend of equivalent positions even ifthey are drawn in different locations from compound to compound or fromformula to formula. For example, in the two pyridine rings (A and B)below, the double bonds are drawn in different locations, however, theyare known to be the same structure and compound:

The present invention includes compounds as well as theirpharmaceutically acceptable salts. Accordingly, the word “or” in thecontext of “a compound or a pharmaceutically acceptable salt thereof” isunderstood to refer to either: 1) a compound alone or a compound and apharmaceutically acceptable salt thereof (alternative), or 2) a compoundand a pharmaceutically acceptable salt thereof (in combination).

Unless indicated otherwise, the nomenclature of substituents that arenot explicitly defined herein are arrived at by naming the terminalportion of the functionality followed by the adjacent functionalitytoward the point of attachment. For example, the substituent“arylalkyloxycarbonyl” refers to the group (aryl)-(alkyl)-O—C(O)—. In aterm such as “—C(R^(x))₂”, it should be understood that the two R^(x)groups can be the same, or they can be different if R^(x) is defined ashaving more than one possible identity. In addition, certainsubstituents are drawn as —R^(x)R^(y), where the “-” indicates a bondadjacent to the parent molecule and R^(y) being the terminal portion ofthe functionality. Similarly, it is understood that the abovedefinitions are not intended to include impermissible substitutionpatterns (e.g., methyl substituted with 5 fluoro groups). Suchimpermissible substitution patterns are well known to the skilledartisan.

In one aspect, there is provided a compound of the formula (I):

wherein:

R₁ is:

wherein

X is selected from the group consisting of NH₂, F and Cl;

R⁵ is selected from the group consisting of H and (C₁-C₁₄) alkyl;

R⁶ is selected from the group consisting of H and —(C═O)—(C₁-C₁₄) alkyl;

R² is selected from the group consisting of (C₁-C₂₄) alkyl;(CH₂)_(n1)—O—(CH₂CH₂O)_(n2)—(C₁-C₁₄ alkyl) where n1 and n2 are integersindependently selected from 1-4-R⁷—NH—(C═O)—R⁸ wherein R⁷ may be(C₁-C₁₄) alkyl and R^(a) may be independently selected from H and(C₁-C₁₄) alkyl; —R⁹—(C₆-C₁₄) aryl, wherein R⁹ is a bond or (C₁-C₆)alkyl; —R¹⁰—(C₃-C₁₄) cycloalkyl, wherein R¹⁰ is a bond or (C₁-C₆) alkyl;—(C₁-C₂₀) alkylene-(C═O)—O—R¹¹ wherein R¹¹ may be selected from H and(C₁-C₂₀)alkyl; and

and;

R³ is selected from the group consisting of H, —(C═O)—(C₁-C₂₄) alkyl;—(C═O)—O—(C₁-C₂₄) alkyl; and C₃-C₁₄ cycloalkyl, or

R² and R³ join together to form a C₃ to C₂₈ cyclic structure; and

with the proviso that when R² is (C₁-C₁₄ alkyl) at least one of R³, R⁵and R⁶ is not H.

In another aspect, the invention provides a compound of the formula(Ia):

wherein:

R₁ is:

wherein:

X is selected from the group consisting of NH₂, F and Cl;

R⁵ is selected from the group consisting of H and (C₁-C₁₄) alkyl;

R⁶ is selected from the group consisting of H and —(C═O)—(C₁-C₁₄) alkyl;

R² is selected from the group consisting of (C₁-C₂₄) alkyl; C₃-C₁₄cycloalkyl; (CH₂)_(n1)—O—(CH₂CH₂O)_(n2)—(C₁-C₁₄ alkyl) where n1 and n2are integers independently selected from 1-4, —R⁷—NH—(C═O)—R⁸ wherein R⁷may be (C₁-C₁₄) alkyl and R^(a) may be independently selected from H and(C₁-C₁₄) alkyl; —R⁹—(C₆-C₁₄) aryl, wherein R⁹ is a bond or (C₁-C₆)alkyl; —R¹⁰—(C₃-C₁₄) cycloalkyl, wherein R¹⁰ is a bond or (C₁-C₆) alkyl;—(C₁-C₂₀) alkylene-(C═O)—O—R¹¹ wherein R¹¹ may be selected from H and(C₁-C₂₀)alkyl; and

and;

R³ is selected from the group consisting of H, —(C═O)—(C₁-C₂₄) alkyl;—(C═O)—O—(C₁-C₂₄) alkyl; and C₃-C₁₄ cycloalkyl; or

R² and R³ join together to form a C₃ to C₂₈ cyclic structure; and

with the proviso that when R² is (C₁-C₁₈ alkyl) at least one of R³, R⁵and R⁶ is not H.

In another aspect, there is provided a compound of the formula (II):

wherein:

R₁ is:

wherein

X is selected from the group consisting of NH₂, F and Cl;

R⁵ is selected from the group consisting of H and (C₁-C₁₄) alkyl;

R⁶ is selected from the group consisting of H and —(C═O)—(C₁-C₁₄) alkyl;

R² is selected from the group consisting of (C₁-C₂₄) alkyl;(CH₂)_(n1)—O—(CH₂CH₂O)_(n2)—(C₁-C₁₄ alkyl) where n1 and n2 are integersindependently selected from 1-4-R⁷—NH—(C═O)—R⁸ wherein R⁷ may be(C₁-C₁₄) alkyl and R^(a) may be independently selected from H and(C₁-C₁₄) alkyl; —R⁹—(C₆-C₁₄) aryl, wherein R⁹ is a bond or (C₁-C₆)alkyl; —R¹⁰—(C₃-C₁₄) cycloalkyl, wherein R¹⁰ is a bond or (C₁-C₆) alkyl;—(C₁-C₂₀) alkylene-(C═O)—O—R¹¹ wherein R¹¹ may be selected from H and(C₁-C₂₀) alkyl; and

and;

R³ is selected from the group consisting of H, —(C═O)—(C₁-C₂₄) alkyl;—(C═O)—O—(C₁-C₂₄) alkyl; and C₃-C₁₄ cycloalkyl; or

R² and R³ join together to form a C₃ to C₂₈ cyclic structure.

In another aspect, the invention provides a compound of the formula(IIa):

wherein:

R₁ is:

wherein:

X is selected from the group consisting of NH₂, F and Cl;

R⁵ is selected from the group consisting of H and (C₁-C₁₄) alkyl;

R⁶ is selected from the group consisting of H and —(C═O)—(C₁-C₁₄) alkyl;

R² is selected from the group consisting of (C₁-C₂₄) alkyl; C₃-C₁₄cycloalkyl; (CH₂)_(n1)—O—(CH₂CH₂O)_(n2)—(C₁-C₁₄ alkyl) where n1 and n2are integers independently selected from 1-4, —R⁷—NH—(C═O)—R⁸ wherein R⁷may be (C₁-C₁₄) alkyl and R^(a) may be independently selected from H and(C₁-C₁₄) alkyl; —R⁹—(C₆-C₁₄) aryl, wherein R⁹ is a bond or (C₁-C₆)alkyl; —R¹⁰—(C₃-C₁₄) cycloalkyl, wherein R¹⁰ is a bond or (C₁-C₆) alkyl;—(C₁-C₂₀) alkylene-(C═O)—O—R¹¹ wherein R¹¹ may be selected from H and(C₁-C₂₀) alkyl; and

and;

R³ is selected from the group consisting of H, —(C═O)—(C₁-C₂₄) alkyl;—(C═O)—O—(C₁-C₂₄) alkyl; and C₃-C₁₄ cycloalkyl; or

R² and R³ join together to form a C₃ to C₂₈ cyclic structure.

In one embodiment of the present invention, there is provided a compoundof the formula (I), (Ia), (II) or (IIa), wherein X is F, and R⁵ and R⁶are each H.

In one embodiment of the present invention, there is provided a compoundof the formula (I), (Ia), (II) or (IIa), wherein X is F, R⁵ is H and R⁶is —(C═O)—(C₁-C₁₄) alkyl

In one embodiment of the present invention, there is provided a compoundof the formula (I), (Ia), (II) or (IIa), wherein R³ is H.

In one embodiment of the present invention, there is provided a compoundof the formula (I), (Ia), (II) or (IIa), wherein R³ is —(C═O)—(C₁-C₂₀)alkyl

In one embodiment of the present invention, there is provided a compoundof the formula (I), (Ia), (II) or (IIa), wherein R² is C₆ aryl.

In one embodiment of the present invention, there is provided a compoundof the formula (I), (Ia), (II) or (IIa), wherein R² is C₆ cycloalkyl.

In one embodiment of the present invention, there is provided a compoundof the formula (I), (Ia), (II) or (IIa), wherein R³ is H, X is F, R⁵ isH, R⁶ is H, and R² is —R⁷—NH—(C═O)—R⁸.

In one embodiment of the present invention, there is provided a compoundof the formula (I), (Ia), (II) or (IIa), wherein R³ is H, X is F, R⁵ isH, R⁶ is H, and R² is —(C₁-C₂₀) alkylene-(C═O)—O—R¹¹

In one embodiment of the present invention, there is provided a compoundof the formula (I), (Ia), (II) or (IIa), wherein R⁹ is a bond.

In one embodiment of the present invention, there is provided a compoundof the formula (I), (Ia), (II) or (IIa), wherein R⁹ is C₁alkyl.

In one embodiment of the present invention, there is provided a compoundof the formula (I), (Ia), (II) or (IIa), wherein R² is—(CH₂)_(n1)—O—(CH₂CH₂O)_(n2)—(C₁-C₁₄alkyl) where n1 and n2 are integersindependently selected from 1-4, more preferably n1 and n2 areindependently selected from 1 and 2. In one preferred embodiment, R² isof the formula —(CH₂)—O—(CH₂CH₂O)_(n2)—(C₁-C₂ alkyl) wherein n2 is 1 or2.

In one embodiment of the present invention, there is provided a compoundof the formula (I), (Ia), (II) or (IIa), wherein R¹⁰ is H.

In one embodiment of the present invention, there is provided a compoundof the formula (I), (Ia), (II) or (IIa), wherein R¹⁰ is C₁ alkyl.

In one preferred embodiment of the present invention, R² is—(CH₂)₈—(C═O)—OH. In one preferred embodiment, R² is —(CH₂)₁₂—(C═O)—OH.In one preferred embodiment, R² is —(CH₂)₁₈—(C═O)—OH.

In one embodiment of the present invention, there is provided a compoundof the formula (I), (Ia), (II) or (IIa), wherein R² is—(CH₂)₃—(C═O)—O—C₁₄ alkyl.

In one embodiment of the present invention, there is provided a compoundof the formula (I), (Ia), (II) or (IIa), wherein R² is of the formula:

In one embodiment of the present invention, there is provided a compoundof the formula (I), (Ia), (II) or (IIa), wherein R² is of the formula:

In one embodiment of the present invention, there is provided a compoundof the formula (I), (Ia), (II) or (IIa), wherein R² is (C₁-C₂₄) alkyland R³ is —(C═O)—(C₁-C₂₀) alkyl

In one embodiment of the present invention, there is provided a compoundof the formula (I), (Ia), (II) or (IIa), wherein R² is (C₁-C₂₄) alkyl,R⁵ is H and R⁶ is —(C═O)—(C₁-C₁₄) alkyl.

In one embodiment of the present invention, there is provided a compoundof the formula (I), (Ia), (II) or (IIa), wherein R³ is H, R⁵ is H, R⁶ isH, and R² is R⁷—NH—(C═O)—R⁸ wherein R⁷ (C₁-C₄) alkyl and R^(a) is(C₁-C₁₄) alkyl.

In one embodiment of the present invention, there is provided a compoundof the formula (I), (Ia), (II) or (IIa), wherein R³ is —(C═O)—(C₁-C₂₄)alkyl, R⁵ is H, R⁶ is H, R² is —(C₁-C₂₄) alkyl and X is F.

In one embodiment of the present invention, there is provided a compoundof the formula (I), (Ia), (II) or (IIa), wherein R³ is —(C═O)—O—(C₁-C₂₄)alkyl, R⁵ is H, R⁶ is H, R² is —(C₁-C₂₄) alkyl and X is F.

In one embodiment of the present invention, there is provided a compoundwherein R² and R³ join together to form a C₃ to C₂₈ cyclic structure.More preferably, in such an embodiment, R³ contains —(C═O)— groupattached at one end to the oxygen substituent of formula (I); inpreferred embodiments, accordingly, R² and R³ join together wherein R²is —(CH₂)_(p)— and R³ is —(C═O)—(CH₂)_(q)— wherein p and q are selectedsuch that a C₃ to C₂₈ cyclic structure is formed. In a most preferredembodiment, p and q are selected such that a C₈ to C₁₈ structure isformed.

The compounds of the present invention may be optionally substituted byone or more substituents. For example, in one embodiment, each of R²,R³, R⁵, R⁶, R⁷, R⁸, R⁹ R¹⁰ and R¹¹ may be independently and optionallysubstituted by one or more (C₁-C₁₄) alkyl, Cl, F, oxo, or (C₁-C₆)alkoxy. In another embodiment, each of R², R³, R⁵, R⁶, R⁷, R⁸, R⁹ R¹⁰and R¹¹ may be independently and optionally substituted by one or more(C₁-C₆) alkyl, Cl, F, oxo, or (C₁-C₆) alkoxy. Preferably, as an example,each of the (C₆-C₁₄) aryl and (C₃-C₁₄) cycloalkyl groups may beoptionally substituted by one or more substituents from (C₁-C₅) alkyl,Cl, F, oxo, or (C₁-C₆) alkoxy. In various preferred embodiments, each ofR², R³, R⁵, R⁶, R⁷, R⁸, R⁹ R¹⁰ and R¹¹, and in particular (C₆-C₁₄) aryl(e.g., (C₆) aryl may be substituted by one or more —C—O—(C═O)—(C₁-C₆alkyl).

In another aspect of the present invention, the invention may encompassvarious individual compounds. As an example, such specific compounds maybe selected from the group consisting of Table 1:

TABLE 1 Example Structure Chemical Name 1

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl cyclohexanecarboxylate 2

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl benzoate 3

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl 2-phenylacetate 4

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl trans-4-(tert-butyl)cyclohexne-1-carboxylate 5

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl trans-4-pentylcyclohexane-1-carboxylate 6

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl 2-(2-methoxyethoxy)acetate 7

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl 2-(2-(2- methoxyethoxy)ethoxy)acetate8

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl 2-(2-butoxyethoxy)acetate 9

10-(((2R,3S,5R)-5-(6-amino-2- fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2- yl)methoxy)-10-oxodecanoic acid 10

14-(((2R,3S,5R)-5-(6-amino-2- fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2- yl)methoxy)-14-oxotetradecanoic acid 11

20-(((2R,3S,5R)-5-(6-amino-2- fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2- yl)methoxy)-20-oxoicosanoic acid 12

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl (1,3-bis(heptanoyloxy)propan-2-yl)glutarate 13

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl (1,3-bis(tetradecanoyloxy)propan-2-yl) glutarate 14

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl tetradecyl glutarate 15

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-3-((ethoxycarbonyl)oxy)-2- ethynyltetrahydrofuran-2-yl)methyl decanoate 16

(2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-2-((heptanoyloxy)methyl)tetra- hydrofuran-3-yl heptanoate 17

(2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-2-((tetradecanoyloxy)methyl)tetra- hydrofuran-3-yl tetradecanoate 18

(2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-((decanoyloxy)methyl)-2- ethynyltetrahydrofuran-3-yl decanoate 19

(2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-2-((pentanoyloxy)methyl)tetra- hydrofuran-3-yl tridecanoate 20

(2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-2-((pentanoyloxy)methyl)tetra- hydrofuran-3-yl heptanoate 21

(2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-2-((pentanoyloxy)methyl)tetra- hydrofuran-3-yl undecanoate 22

(2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-2-((pentanoyloxy)methyl)tetra- hydrofuran-3-yl nonanoate 23

(2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-2-((pentanoyloxy)methyl)tetra- hydrofuran-3-yl pentanoate 24

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-(propionyloxy)tetrahydrofuran-2-yl) methyl pentanoate 25

((2R,3S,5R)-5-(6-butyramido-2- fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl) methyl pentanoate 26

((2R,3S,5R)-2-ethynyl-5-(2-fluoro- 6-octanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methyl pentanoate 27

((2R,3S,5R)-2-ethynyl-5-(2-fluoro- 6-tetradecanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2- yl)methyl pentanoate 28

((2R,3S,5R)-5-(6-dodecanamido-2- fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl pentanoate 29

(2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-2-(((2-propylpentanoyl)oxy)methyl) tetrahydrofuran-3-yl dodecanoate 30

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-3-(butyryloxy)-2-ethynyltetrahydrofuran-2-yl)methyl 2-propylpentanoate 31

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-3-(butyryloxy)-2-ethynyltetrahydrofuran-2-yl)methyl decanoate 32

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-(heptanoyloxy)tetrahydrofuran-2- yl)methyl benzoate 33

(2R,3aS,20aR)-2-(6-amino-2- fluoro-9H-purin-9-yl)-20a-ethynylhexadecahydro-2H-furo[3,2- b][1,5]dioxacyclononadecine-5,18-dione 34

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl) methyl heptanoylglycinate 35

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl tetradecanoylglycinate 36

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl heptanoyl-L-alaninate 37

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-3-(butyryloxy)-2-ethynyltetrahydrofuran-2-yl)methyl tetradecanoate 38

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy) tetrahydrofuran-2-yl) methyl tetradecanoate 39

(2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-2-(((2-propylpentanoyl)oxy)methyl) tetrahydrofuran-3-yl heptanoate 40

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl tetradecanoyl-L-alaninate 41

(2R,3S,5R)-2-(acetoxymethyl)-5-(6- amino-2-fluoro-9H-purin-9-yl)-2-ethynyltetrahydrofuran-3-yl heptanoate 42

((2R,3S,5R)-5-(6-decanamido-2- fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl) methyl heptanoate 43

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy) tetrahydrofuran-2-yl)methyl heptanoate 44

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy) tetrahydrofuran-2-yl)methyl decanoate 45

((2R,3S,5R)-2-ethynyl-5-(2-fluoro- 6-tetradecanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2- yl)methyl benzoate 46

((2R,3S,5R)-5-(6-decanamido-2- fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl benzoate 47

((2R,3S,5R)-2-ethynyl-5-(2-fluoro- 6-tetradecanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2- yl)methyl heptanoate 48

((2R,3S,5R)-2-ethynyl-5-(2-fluoro- 6-tetradecanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2- yl)methyl decanoate 49

((2R,3S,5R)-2-ethynyl-5-(2-fluoro- 6-tetradecanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2- yl)methyl 2-propylpentanoate 50

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl 3-(2-acetoxy-4,6-dimethylphenyl)-3-methylbutanoate 51

(2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-2-((heptanoyloxy)methyl)tetra- hydrofuran-3-yl tetradecanoate 52

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-(((tridecyloxy)carbonyl)oxy)tetra- hydrofuran-2-yl)methyl heptanoate 53

(2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-2-((propionyloxy)methyl)tetra- hydrofuran-3-yl stearate 54

((2R,3S,5R)-2-ethynyl-5-(2-fluoro- 6-heptanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methyl 2-propylpentanoate 55

((2R,3S,5R)-2-ethynyl-5-(2-fluoro- 6-tetradecanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2- yl)methyl propionate 56

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-3-((ethoxycarbonyl)oxy)-2- ethynyltetrahydrofuran-2-yl)methyltetradecanoate 57

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl (1,3-bis(isobutyryloxy)propan-2-yl)succinate 58

(2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-((dodecanoyloxy)methyl)-2- ethynyltetrahydrofuran-3-yl dodecanoate 59

(2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-2-((palmitoyloxy)methyl)tetra- hydrofuran-3-yl palmitate 60

(2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-2-((icosanoyloxy)methyl)tetra- hydrofuran-3-yl icosanoate 61

(2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-2-((undecanoyloxy)methyl)tetra- hydrofuran-3-yl undecanoate 62

(2R,3aS,26aR)-2-(6-amino-2- fluoro-9H-purin-9-yl)-26a-ethynyldocosahydro-2H-furo[3,2- b][1,5]dioxacyclopentacosine-5,24- dione63

(2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-2-((hexanoyloxy)methyl)tetra- hydrofuran-3-yl hexanoate 64

(2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-2-((nonanoyloxy)methyl)tetra- hydrofuran-3-yl nonanoate 65

(2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-2-((octanoyloxy)methyl)tetra- hydrofuran-3-yl octanoate 66

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl 3-(2,4-dimethyl-6-(pivaloyloxy)phenyl)-3- methylbutanoate 67

2-(4-(((2R,3S,5R)-5-(6-amino-2- fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2- yl)methoxy)-2-methyl-4-oxobutan-2-yl)-3,5-dimethylphenyl decanoate 68

((2R,3S,5R)-5-(6-amino-2-fluoro- 9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl (1,3-bis(heptanoyloxy)propan-2-yl)succinate

and pharmaceutically acceptable salts thereof.

In one embodiment, the present invention encompasses each individualcompound listed in the above Table 1, or a pharmaceutically acceptablesalt thereof.

In various embodiments, prodrugs of any of the compounds of formula (I),(Ia), (II) or (IIa) set forth herein are also within the scope of thepresent invention.

In accordance with one embodiment of the present invention, there isprovided a pharmaceutical composition comprising a compound of Formulas(I), (Ia), (II) or (IIa) or a pharmaceutically acceptable salt thereof,and a pharmaceutically acceptable excipient. In a further embodiment, acompound is present in amorphous form. In a further embodiment, thecompound is present in crystalline form. In various preferredembodiments, in particular, the compounds of the structure of Examples18, 38 and 58 may be each present in crystalline form as well asamorphous form. In a further embodiment, the pharmaceutical compositionis in a tablet form. In a further embodiment, the pharmaceuticalcomposition is in parenteral form. In a further embodiment, the compoundis present as a spray dried dispersion.

In accordance with one embodiment of the present invention, there isprovided a method of treating an HIV infection in a subject comprisingadministering to the subject a compound of Formulas (I), (Ia), (II) or(IIa) or a pharmaceutically acceptable salt thereof.

In accordance with one embodiment of the present invention, there isprovided a method of treating an HIV infection in a subject comprisingadministering to the subject a pharmaceutical composition as describedherein.

In accordance with one embodiment of the present invention, there isprovided a method of preventing an HIV infection in a subject at riskfor developing an HIV infection, comprising administering to the subjecta compound of Formulas (I), (Ia), (II) or (IIa) or a pharmaceuticallyacceptable salt thereof.

In accordance with one embodiment of the present invention, there isprovided the use of a compound of Formula (I), (Ia), (II) or (IIa) inthe manufacture of a medicament for treating an HIV infection.

In accordance with one embodiment of the present invention, there isprovided the use of a compound of Formula (I), (Ia), (II) or (IIa) inthe manufacture of a medicament for preventing an HIV infection.

In accordance with one embodiment of the present invention, there isprovided a compound according to Formula (I), (Ia), (II) or (IIa) foruse in treating an HIV infection.

In accordance with one embodiment of the present invention, there isprovided a compound according to Formula (I), (Ia), (II) or (IIa) foruse in preventing an HIV infection.

In accordance with one embodiment of the present invention, there isprovided a method of preventing an HIV infection in a subject at riskfor developing an HIV infection, comprising administering to the subjecta pharmaceutical composition as described herein.

Furthermore, the compounds of the invention can exist in particulargeometric or stereoisomeric forms. The invention contemplates all suchcompounds, including cis- and trans-isomers, (−)- and (+)-enantiomers,(R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, theracemic mixtures thereof, and other mixtures thereof, such asenantiomerically or diastereomerically enriched mixtures, as fallingwithin the scope of the invention. Additional asymmetric carbon atomscan be present in a substituent such as an alkyl group. All suchisomers, as well as mixtures thereof, are intended to be included inthis invention.

Optically active (R)- and (S)-isomers and d and l isomers can beprepared using chiral synthons or chiral reagents, or resolved usingconventional techniques. If, for instance, a particular enantiomer of acompound of the present invention is desired, it can be prepared byasymmetric synthesis, or by derivatization with a chiral auxiliary,where the resulting diastereomeric mixture is separated and theauxiliary group cleaved to provide the pure desired enantiomers.Alternatively, where the molecule contains a basic functional group,such as an amino group, or an acidic functional group, such as acarboxyl group, diastereomeric salts can be formed with an appropriateoptically active acid or base, followed by resolution of thediastereomers thus formed by fractional crystallization orchromatographic means known in the art, and subsequent recovery of thepure enantiomers. In addition, separation of enantiomers anddiastereomers is frequently accomplished using chromatography employingchiral, stationary phases, optionally in combination with chemicalderivatization (e.g., formation of carbamates from amines).

In another embodiment of the invention, there is provided a compound ofFormula (I), (Ia), (II) or (IIa) wherein the compound or salt of thecompound is used in the manufacture of a medicament for use in thetreatment of an HIV infection in a human.

In another embodiment of the invention, there is provided a compound ofFormula (I), (Ia), (II) or (IIa) wherein the compound or salt of thecompound is used in the manufacture of a medicament for use in theprevention of an HIV infection in a human.

In one embodiment, the pharmaceutical formulation containing a compoundof Formula (I), (Ia), (II) or (IIa) or a salt thereof is a formulationadapted for parenteral administration. In another embodiment, theformulation is a long-acting parenteral formulation. In a furtherembodiment, the formulation is a nano-particle formulation.

The compounds of the present invention and their salts, solvates, orother pharmaceutically acceptable derivatives thereof, may be employedalone or in combination with other therapeutic agents. Therefore, inother embodiments, the methods of treating and/or preventing an HIVinfection in a subject may in addition to administration of a compoundof Formula (I), (Ia), (II) or (IIa) further comprise administration ofone or more additional pharmaceutical agents active against HIV.

In such embodiments, the one or more additional agents active againstHIV is selected from the group consisting of zidovudine, didanosine,lamivudine, zalcitabine, abacavir, stavudine, adefovir, adefovirdipivoxil, fozivudine, todoxil, emtricitabine, alovudine, amdoxovir,elvucitabine, nevirapine, delavirdine, efavirenz, loviride, immunocal,oltipraz, capravirine, lersivirine, GSK2248761, TMC-278, TMC-125,etravirine, saquinavir, ritonavir, indinavir, nelfinavir, amprenavir,fosamprenavir, brecanavir, darunavir, atazanavir, tipranavir, palinavir,lasinavir, enfuvirtide, T-20, T-1249, PRO-542, PRO-140, TNX-355,BMS-806, BMS-663068 and BMS-626529, 5-Helix, raltegravir, elvitegravir,dolutegravir, cabotegravir, bictegravir, vicriviroc (Sch-C), Sch-D,TAK779, maraviroc, TAK449, didanosine, tenofovir, lopinavir, anddarunavir.

As such, the compounds of the present invention of Formulas (I), (Ia),(II) or (IIa) and any other pharmaceutically active agent(s) may beadministered together or separately and, when administered separately,administration may occur simultaneously or sequentially, in any order.The amounts of the compounds of Formula (I), (Ia), (II) or (IIa) of thepresent invention and the other pharmaceutically active agent(s) and therelative timings of administration will be selected in order to achievethe desired combined therapeutic effect. The administration incombination of a compound of the present invention of Formula (I), (Ia),(II) or (IIa) and salts, solvates, or other pharmaceutically acceptablederivatives thereof with other treatment agents may be in combination byadministration concomitantly in: (1) a unitary pharmaceuticalcomposition including both compounds; or (2) separate pharmaceuticalcompositions each including one of the compounds. Alternatively, thecombination may be administered separately in a sequential mannerwherein one treatment agent is administered first and the other secondor vice versa. Such sequential administration may be close in time orremote in time. The amounts of the compound(s) of Formula (I), (Ia),(II) or (IIa) or salts thereof and the other pharmaceutically activeagent(s) and the relative timings of administration will be selected inorder to achieve the desired combined therapeutic effect.

In addition, the compounds of the present invention of Formula (I),(Ia), (II) or (IIa) may be used in combination with one or more otheragents that may be useful in the prevention or treatment of HIV.Examples of such agents include:

Nucleotide reverse transcriptase inhibitors such as zidovudine,didanosine, lamivudine, zalcitabine, abacavir, stavudine, adefovir,adefovir dipivoxil, fozivudine, todoxil, emtricitabine, alovudine,amdoxovir, elvucitabine, and similar agents;Non-nucleotide reverse transcriptase inhibitors (including an agenthaving anti-oxidation activity such as immunocal, oltipraz, etc.) suchas nevirapine, delavirdine, efavirenz, loviride, immunocal, oltipraz,capravirine, lersivirine, doravirine, GSK2248761, TMC-278, TMC-125,etravirine, and similar agents;Protease inhibitors such as saquinavir, ritonavir, indinavir,nelfinavir, amprenavir, fosamprenavir, brecanavir, darunavir,atazanavir, tipranavir, palinavir, lasinavir, and similar agents;Entry, attachment and fusion inhibitors such as enfuvirtide (T-20),T-1249, PRO-542, PRO-140, TNX-355, BMS-806, BMS-663068 (Fostemsavir),BMS-626529 (Temsavir), 5-Helix and similar agents;Integrase inhibitors such as raltegravir, elvitegravir, dolutegravir,bictegravir, cabotegravir and similar agents;Maturation inhibitors such as PA-344 and PA-457, and similar agents; andCXCR4 and/or CCR5 inhibitors such as vicriviroc (Sch-C), Sch-D, TAK779,maraviroc (UK 427,857), TAK449, as well as those disclosed in WO02/74769, PCT/US03/39644, PCT/US03/39975, PCT/USO3/39619,PCT/USO3/39618, PCT/US03/39740, and PCT/US03/39732, and similar agents.

CAPSID inhibitors such GS-6207, and similar agents.

Further examples where the compounds of the present invention may beused in combination with one or more agents useful in the prevention ortreatment of HIV are found in Table 2.

TABLE 2 Brand FDA Approval Name Generic Name Manufacturer NucleosideReverse Transcriptase Inhibitors (NRTIs) 1987 Retrovir zidovudine,GlaxoSmithKline azidothymidine, AZT, ZDV 1991 Videx didanosine,Bristol-Myers dideoxyinosine, Squibb ddI 1992 Hivid zalcitabine, Rochedideoxycytidine, Pharmaceuticals ddC 1994 Zerit stavudine, d4TBristol-Myers Squibb 1995 Epivir lamivudine, 3TC GlaxoSmithKline 1997Combivir lamivudine + GlaxoSmithKline zidovudine 1998 Ziagen abacavirsulfate, GlaxoSmithKline ABC 2000 Trizivir abacavir + GlaxoSmithKlinelamivudine + zidovudine 2000 Videx EC enteric coated Bristol-Myersdidanosine, ddI Squibb EC 2001 Viread tenofovir disoproxil GileadSciences fumarate, TDF 2003 Emtriva emtricitabine, FTC Gilead Sciences2004 Epzicom abacavir + GlaxoSmithKline lamivudine 2004 Truvadaemtricitabine + Gilead Sciences tenofovir disoproxil fumarateNon-Nucleosides Reverse Transcriptase Inhibitors (NNRTIs) 1996 Viramunenevirapine, NVP Boehringer Ingelheim 1997 Rescriptor delavirdine, DLVPfizer 1998 Sustiva efavirenz, EFV Bristol-Myers Squibb 2008 IntelenceEtravirine Tibotec Therapeutics 2011 Edurant Rilpivirine TibotecTherapeutics 2018 Pifeltro Doravirine Merck Protease Inhibitors (PIs)1995 Invirase saquinavir Roche mesylate, SQV Pharmaceuticals 1996 Norvirritonavir, RTV Abbott Laboratories 1996 Crixivan indinavir, IDV Merck1997 Viracept nelfinavir Pfizer mesylate, NFV 1997 Fortovase saquinavir(no Roche longer marketed) Pharmaceuticals 1999 Agenerase amprenavir,APV GlaxoSmithKline 2000 Kaletra lopinavir + Abbott ritonavir, LPV/RTVLaboratories 2003 Reyataz atazanavir sulfate, Bristol-Myers ATV Sguibb2003 Lexiva fosamprenavir GlaxoSmithKline calcium, FOS-APV 2005 Aptivustripranavir, TPV Boehringer Ingelheim 2006 Prezista Darunavir TibotecTherapeutics Fusion Inhibitors 2003 Fuzeon Enfuvirtide, T-20 RochePharmaceuticals & Trimeris Entry Inhibitors 2007 Selzentry MaravirocPfizer Integrase Inhibitors 2007 Isentress Raltegravir Merck 2013Tivicay Dolutegravir ViiV Healthcare 2018 Bictegravir Gilead Sciences —— Cabotegravir ViiV Healthcare Capsid Inhibitors — GS-6207 GileadSciences

The scope of combinations of compounds of this invention with HIV agentsis not limited to those mentioned above, but includes in principle anycombination with any pharmaceutical composition useful for the treatmentand/or prevention of HIV. As noted, in such combinations the compoundsof the present invention and other HIV agents may be administeredseparately or in conjunction. In addition, one agent may be prior to,concurrent to, or subsequent to the administration of other agent(s).

The present invention may be used in combination with one or more agentsuseful as pharmacological enhancers as well as with or withoutadditional compounds for the prevention or treatment of HIV. Examples ofsuch pharmacological enhancers (or pharmakinetic boosters) include, butare not limited to, ritonavir, GS-9350, and SPI-452. Ritonavir is10-hydroxy-2-methyl-5-(1-methyethyl)-1-1[2-(1-methylethyl)-4-thiazolyl]-3,6-dioxo-8,11-bis(phenylmethyl)-2,4,7,12-tetraazatridecan-13-oicacid, 5-thiazolylmethyl ester, [5S-(5S*,8R*,10R*,11R*)] and is availablefrom Abbott Laboratories of Abbott park, Illinois, as Norvir. Ritonaviris an HIV protease inhibitor indicated with other antiretroviral agentsfor the treatment of HIV infection. Ritonavir also inhibits P450mediated drug metabolism as well as the P-gycoprotein (Pgp) celltransport system, thereby resulting in increased concentrations ofactive compound within the organism.

GS-9350 is a compound being developed by Gilead Sciences of Foster CityCalif. as a pharmacological enhancer.SPI-452 is a compound being developed by Sequoia Pharmaceuticals ofGaithersburg, Md., as a pharmacological enhancer.

In one embodiment of the present invention, a compound of Formula (I),(Ia), (II) or (IIa) is used in combination with ritonavir. In oneembodiment, the combination is an oral fixed dose combination. Inanother embodiment, the compound of Formula (I), (Ia), (II) or (IIa) isformulated as a long acting parenteral injection and ritonavir isformulated as an oral composition. In one embodiment, a kit containingthe compound of Formula (I), (Ia), (II) or (IIa) is formulated as a longacting parenteral injection and ritonavir formulated as an oralcomposition. In another embodiment, the compound of Formula (I), (Ia),(II) or (IIa) is formulated as a long acting parenteral injection andritonavir is formulated as an injectable composition. In one embodiment,a kit containing the compound of Formula (I), (Ia), (II) or (IIa) isformulated as a long acting parenteral injection and ritonavirformulated as an injectable composition.

In another embodiment of the present invention, a compound of Formula(I), (Ia), (II) or (IIa) is used in combination with GS-9350. In oneembodiment, the combination is an oral fixed dose combination. Inanother embodiment, the compound of Formula (I), (Ia), (II) or (IIa) isformulated as a long acting parenteral injection and GS-9350 isformulated as an oral composition. In one embodiment, there is provideda kit containing the compound of Formula (I), (Ia), (II) or (IIa) isformulated as a long acting parenteral injection and GS-9350 formulatedas an oral composition. In another embodiment, the compound of Formula(I), (Ia), (II) or (IIa) is formulated as a long acting parenteralinjection and GS-9350 is formulated as an injectable composition. In oneembodiment, is a kit containing the compound of Formula (I), (Ia), (II)or (IIa) is formulated as a long acting parenteral injection and GS-9350formulated as an injectable composition.

In one embodiment of the present invention, a compound of Formula (I),(Ia), (II) or (IIa) is used in combination with SPI-452. In oneembodiment, the combination is an oral fixed dose combination. Inanother embodiment, the compound of Formula (I), (Ia), (II) or (IIa) isformulated as a long acting parenteral injection and SPI-452 isformulated as an oral composition. In one embodiment, there is provideda kit containing the compound of Formula (I) formulated as a long actingparenteral injection and SPI-452 formulated as an oral composition. Inanother embodiment, the compound of Formula (I), (Ia), (II) or (IIa) isformulated as a long acting parenteral injection and SPI-452 isformulated as an injectable composition. In one embodiment, there isprovided a kit containing the compound of Formula (I), (Ia), (II) or(IIa) formulated as a long acting parenteral injection and SPI-452formulated as an injectable composition.

In one embodiment of the present invention, a compound of Formula (I),(Ia), (II) or (IIa) is used in combination with compounds which arefound in previously filed PCT/CN2011/0013021, which is hereinincorporated by reference.

The above other therapeutic agents, when employed in combination withthe chemical entities described herein, may be used, for example, inthose amounts indicated in the Physicians' Desk Reference (PDR) or asotherwise determined by one of ordinary skill in the art.

In another embodiment of the invention, there is provided a method fortreating a viral infection in a mammal mediated at least in part by avirus in the retrovirus family of viruses which method comprisesadministering to a mammal, that has been diagnosed with said viralinfection or is at risk of developing said viral infection, a compoundof Formula (I), (Ia), (II) or (IIa).

In another embodiment of the invention, there is provided a method fortreating a viral infection in a mammal mediated at least in part by avirus in the retrovirus family of viruses which method comprisesadministering to a mammal, that has been diagnosed with said viralinfection or is at risk of developing said viral infection, a compoundof Formula (I), (Ia), (II) or (IIa), wherein said virus is an HIV virus.In some embodiments, the HIV virus is the HIV-1 virus.

In another embodiment of the invention, there is provided a method fortreating a viral infection in a mammal mediated at least in part by avirus in the retrovirus family of viruses which method comprisesadministering to a mammal, that has been diagnosed with said viralinfection or is at risk of developing said viral infection, a compoundof Formula (I), (Ia), (II) or (IIa) further comprising administration ofa therapeutically effective amount of one or more agents active againstan HIV virus.

In another embodiment of the invention, there is provided a method fortreating a viral infection in a mammal mediated at least in part by avirus in the retrovirus family of viruses which method comprisesadministering to a mammal, that has been diagnosed with said viralinfection or is at risk of developing said viral infection, a compoundof Formula (I), (Ia), (II) or (IIa), further comprising administrationof a therapeutically effective amount of one or more agents activeagainst the HIV virus, wherein said agent active against HIV virus isselected from Nucleotide reverse transcriptase inhibitors;Non-nucleotide reverse transcriptase inhibitors; Protease inhibitors;Entry, attachment and fusion inhibitors; Integrase inhibitors;Maturation inhibitors; CAPSID inhibitors, CXCR4 inhibitors; and CCR5inhibitors.

In another embodiment of the invention, there is provided a method forpreventing a viral infection in a mammal mediated at least in part by avirus in the retrovirus family of viruses which method comprisesadministering to a mammal, that has been diagnosed with said viralinfection or is at risk of developing said viral infection, a compoundof Formula (I), (Ia), (II) or (IIa).

In another embodiment of the invention, there is provided a method forpreventing a viral infection in a mammal mediated at least in part by avirus in the retrovirus family of viruses which method comprisesadministering to a mammal, that has been diagnosed with said viralinfection or is at risk of developing said viral infection, a compoundof Formula (I), (Ia), (II) or (IIa), wherein said virus is an HIV virus.In some embodiments, the HIV virus is the HIV-1 virus.

In another embodiment of the invention, there is provided a method forpreventing a viral infection in a mammal mediated at least in part by avirus in the retrovirus family of viruses which method comprisesadministering to a mammal, that has been diagnosed with said viralinfection or is at risk of developing said viral infection, a compoundof Formula (I), (Ia), (II) or (IIa), further comprising administrationof a therapeutically effective amount of one or more agents activeagainst an HIV virus.

In another embodiment of the invention, there is provided a method forpreventing a viral infection in a mammal mediated at least in part by avirus in the retrovirus family of viruses which method comprisesadministering to a mammal, that has been diagnosed with said viralinfection or is at risk of developing said viral infection, a compoundof Formula (I), (Ia), (II) or (IIa) further comprising administration ofa therapeutically effective amount of one or more agents active againstthe HIV virus, wherein said agent active against HIV virus is selectedfrom Nucleotide reverse transcriptase inhibitors; Non-nucleotide reversetranscriptase inhibitors; Protease inhibitors; Entry, attachment andfusion inhibitors; Integrase inhibitors; Maturation inhibitors; CAPSIDinhibitors, CXCR4 inhibitors; and CCR5 inhibitors.

In further embodiments, the compound of the present invention of Formula(I), (Ia), (II) or (IIa) or a pharmaceutically acceptable salt thereof,is selected from the group of compounds set forth in Table 1 above.

The compounds of Table 1 were synthesized according to the SyntheticMethods, General Schemes, and the Examples described below.

In another embodiment, there is provided a pharmaceutical compositioncomprising a pharmaceutically acceptable diluent and a therapeuticallyeffective amount of a compound of Formula (I), (Ia), (II) or (IIa) or apharmaceutically acceptable salt thereof.

In certain embodiments, the compound(s) of the present invention, or apharmaceutically acceptable salt thereof, is chosen from the compoundsset forth in Table 1.

The compounds of the present invention can be supplied in the form of apharmaceutically acceptable salt. The term “pharmaceutically acceptablesalt” may refer to salts prepared from pharmaceutically acceptableinorganic and organic acids and bases. Accordingly, the word “or” in thecontext of “a compound or a pharmaceutically acceptable salt thereof” isunderstood to refer to either a compound or a pharmaceuticallyacceptable salt thereof (alternative), or a compound and apharmaceutically acceptable salt thereof (in combination).

The compounds of Formula (I), (Ia), (II) or (IIa) of the invention mayexist in both unsolvated and solvated forms. The term ‘solvate’comprises the compound of the invention and one or more pharmaceuticallyacceptable solvent molecules, for example, ethanol. The term ‘hydrate’is employed when said solvent is water. Pharmaceutically acceptablesolvates include hydrates and other solvates wherein the solvent ofcrystallization may be isotopically substituted, e.g. D₂O, d₆-acetone,d₆-DMSO.

Compounds of Formula (I), (Ia), (II) or (IIa) containing one or moreasymmetric carbon atoms can exist as two or more stereoisomers. Where acompound of Formula (I), (Ia), (II) or (IIa) contains an alkenyl oralkenylene group or a cycloalkyl group, geometric cis/trans (or Z/E)isomers are possible. Where the compound contains, for example, a ketoor oxime group or an aromatic moiety, tautomeric isomerism(‘tautomerism’) can occur. It follows that a single compound may exhibitmore than one type of isomerism.

Included within the scope of the claimed compounds present invention areall stereoisomers, geometric isomers and tautomeric forms of thecompounds of Formula (I), (Ia), (II) or (IIa), including compoundsexhibiting more than one type of isomerism, and mixtures of one or morethereof. Also included are acid addition or base salts wherein thecounterion is optically active, for example, D-lactate or L-lysine, orracemic, for example, DL-tartrate or DL-arginine.

Cis/trans isomers may be separated by conventional techniques well knownto those skilled in the art, for example, chromatography and fractionalcrystallisation.

Conventional techniques for the preparation/isolation of individualenantiomers include chiral synthesis from a suitable optically pureprecursor or resolution of the racemate (or the racemate of a salt orderivative) using, for example, chiral high pressure liquidchromatography (HPLC), Supercritical fluid chromatography (SFC).

Alternatively, the racemate (or a racemic precursor) may be reacted witha suitable optically active compound, for example, an alcohol, or, inthe case where the compound of Formula (I) contains an acidic or basicmoiety, an acid or base such as tartaric acid or 1-phenylethylamine. Theresulting diastereomeric mixture may be separated by chromatographyand/or fractional crystallization and one or both of thediastereoisomers converted to the corresponding pure enantiomer(s) bymeans well known to a skilled person.

Chiral compounds of the invention (and chiral precursors thereof) may beobtained in enantiomerically-enriched form using chromatography,typically HPLC or SFC, on a resin with an asymmetric stationary phaseand with a mobile phase consisting of a hydrocarbon, typically heptaneor hexane, containing from 0 to 50% isopropanol, typically from 2 to20%, and from 0 to 5% of an alkylamine, typically 0.1% diethylamine.Concentration of the eluate affords the enriched mixture.

Mixtures of stereoisomers may be separated by conventional techniquesknown to those skilled in the art. [see, for example, “Stereochemistryof Organic Compounds” by E L Eliel (Wiley, New York, 1994).]

The present invention includes all pharmaceutically acceptableisotopically-labelled compounds of Formula (I), (Ia), (II) or (IIa)wherein one or more atoms are replaced by atoms having the same atomicnumber, but an atomic mass or mass number different from the atomic massor mass number usually found in nature.

Examples of isotopes suitable for inclusion in the compounds of theinvention include isotopes of hydrogen, such as ²H and ³H, carbon, suchas ¹¹C, ¹³C and ¹⁴C, chlorine, such as ³⁶Cl, fluorine, such as ¹⁸F,iodine, such as ¹²³I and ¹²⁵I, nitrogen, such as ¹³N and ¹⁵N, oxygen,such as ¹⁵O, ¹⁷O and ¹⁸O, phosphorus, such as ³²P, and sulphur, such as³⁵S.

Certain isotopically-labelled compounds of Formula (I), (Ia), (II) or(IIa), for example, those incorporating a radioactive isotope, areuseful in drug and/or substrate tissue distribution studies. Theradioactive isotopes tritium, i.e. ³H, and carbon-14, i.e. ¹⁴C, areparticularly useful for this purpose in view of their ease ofincorporation and ready means of detection. Substitution with heavierisotopes such as deuterium, i.e. ²H, may afford certain therapeuticadvantages resulting from greater metabolic stability, for example,increased in vivo half-life or reduced dosage requirements, and hencemay be preferred in some circumstances.

Isotopically-labelled compounds of Formula (I), (Ia), (II) or (IIa) cangenerally be prepared by conventional techniques known to those skilledin the art or by processes analogous to those described herein using anappropriate isotopically-labelled reagent in place of the non-labelledreagent previously employed.

The compounds of the present invention may be administered as prodrugs.Thus, certain derivatives of compounds of Formula (I), (Ia), (II) or(IIa), which may have little or no pharmacological activity themselvescan, when administered into or onto the body, be converted intocompounds of Formula (I), (Ia), (II) or (IIa) as ‘prodrugs’. One exampleof a compound that such prodrugs may encompass is4′-ethylnyl-2-fluoro-2′-dooxyadenosine (EFdA) disclosed e.g., in U.S.Pat. No. 7,339,053. The compounds of the present invention may beadministered as prodrugs. In one embodiment, the compounds of theinvention are prodrugs of 4′-ethynyl-2-fluoro-2′-deoxyadenosine (EFdA)disclosed e.g., in U.S. Pat. No. 7,339,053, which is a nucleosidereverse transcriptase inhibitor of the formula:

The prodrugs are useful in that they are capable of modulatingphysicochemical properties, facilitating multiple dosing paradigms andimproving pharmacokinetic and/or pharmacodynamic profiles of the activeparent (EfdA). For example, the prodrugs may facilitate long-actingparenteral dosing modalities, and/or improvements in antiviralpersistence profiles as compared to EFdA.

Administration of the chemical entities and combinations of entitiesdescribed herein can be via any of the accepted modes of administrationfor agents that serve similar utilities including, but not limited to,orally, sublingually, subcutaneously, intravenously, intranasally,topically, transdermally, intraperitoneally, intramuscularly,intrapulmonarilly, vaginally, rectally, or intraocularly. In someembodiments, oral or parenteral administration is used. Examples ofdosing include, without limitation, once every seven days for oral, onceevery eight weeks for intramuscular, or once every six months forsubcutaneous.

Pharmaceutical compositions or formulations include solid, semi-solid,liquid and aerosol dosage forms, such as, e.g., tablets, capsules,powders, liquids, suspensions, suppositories, aerosols or the like. Thechemical entities can also be administered in sustained or controlledrelease dosage forms, including depot injections, osmotic pumps, pills,transdermal (including electrotransport) patches, and the like, forprolonged and/or timed, pulsed administration at a predetermined rate.In certain embodiments, the compositions are provided in unit dosageforms suitable for single administration of a precise dose.

The chemical entities described herein can be administered either aloneor more typically in combination with a conventional pharmaceuticalcarrier, excipient or the like (e.g., mannitol, lactose, starch,magnesium stearate, sodium saccharine, talcum, cellulose, sodiumcrosscarmellose, glucose, gelatin, sucrose, magnesium carbonate, and thelike). If desired, the pharmaceutical composition can also contain minoramounts of nontoxic auxiliary substances such as wetting agents,emulsifying agents, solubilizing agents, pH buffering agents and thelike (e.g., sodium acetate, sodium citrate, cyclodextrine derivatives,sorbitan monolaurate, triethanolamine acetate, triethanolamine oleate,and the like). Generally, depending on the intended mode ofadministration, the pharmaceutical composition will contain about 0.005%to 95%; in certain embodiments, about 0.5% to 50% by weight of achemical entity. Actual methods of preparing such dosage forms areknown, or will be apparent, to those skilled in this art; for example,see Remington's Pharmaceutical Sciences, Mack Publishing Company,Easton, Pa.

In certain embodiments, the compositions will take the form of a pill ortablet and thus the composition will contain, along with the activeingredient, a diluent such as lactose, sucrose, dicalcium phosphate, orthe like; a lubricant such as magnesium stearate or the like; and abinder such as starch, gum acacia, polyvinylpyrrolidine, gelatin,cellulose, cellulose derivatives or the like. In another solid dosageform, a powder, marume, solution or suspension (e.g., in propylenecarbonate, vegetable oils or triglycerides) is encapsulated in a gelatincapsule.

Liquid pharmaceutically administrable compositions can, for example, beprepared by dissolving, dispersing, etc. at least one chemical entityand optional pharmaceutical adjuvants in a carrier (e.g., water, saline,aqueous dextrose, glycerol, glycols, ethanol or the like) to form asolution or suspension. Injectables can be prepared in conventionalforms, either as liquid solutions or suspensions, as emulsions, or insolid forms suitable for dissolution or suspension in liquid prior toinjection. The percentage of chemical entities contained in suchparenteral compositions is highly dependent on the specific naturethereof, as well as the activity of the chemical entities and the needsof the subject. However, percentages of active ingredient of 0.01% to10% in solution are employable, and will be higher if the composition isa solid which will be subsequently diluted to the above percentages. Incertain embodiments, the composition may comprise from about 0.2 to 2%of the active agent in solution.

Pharmaceutical compositions of the chemical entities described hereinmay also be administered to the respiratory tract as an aerosol orsolution for a nebulizer, or as a microfine powder for insufflation,alone or in combination with an inert carrier such as lactose. In such acase, the particles of the pharmaceutical composition have diameters ofless than 50 microns, in certain embodiments, less than 10 microns.

In general, the chemical entities provided will be administered in atherapeutically effective amount by any of the accepted modes ofadministration for agents that serve similar utilities. The actualamount of the chemical entity, i.e., the active ingredient, will dependupon numerous factors such as the severity of the disease to be treated,the age and relative health of the subject, the potency of the chemicalentity used the route and form of administration, and other factors. Thedrug can be administered more than once a day, such as once or twice aday.

In general, the chemical entities will be administered as pharmaceuticalcompositions by any one of the following routes: oral, systemic (e.g.,transdermal, intranasal or by suppository), or parenteral (e.g.,intramuscular, intravenous or subcutaneous) administration. In certainembodiments, oral administration with a convenient daily dosage regimenthat can be adjusted according to the degree of affliction may be used.Compositions can take the form of tablets, pills, capsules, semisolids,powders, sustained release formulations, solutions, suspensions,elixirs, aerosols, or any other appropriate compositions. Another mannerfor administering the provided chemical entities is inhalation.

The choice of formulation depends on various factors such as the mode ofdrug administration and bioavailability of the drug substance. Fordelivery via inhalation the chemical entity can be formulated as liquidsolution, suspensions, aerosol propellants or dry powder and loaded intoa suitable dispenser for administration. There are several types ofpharmaceutical inhalation devices-nebulizer inhalers, metered doseinhalers (MDI) and dry powder inhalers (DPI). Nebulizer devices producea stream of high velocity air that causes the therapeutic agents (whichare formulated in a liquid form) to spray as a mist that is carried intothe patient's respiratory tract. MDIs typically are formulation packagedwith a compressed gas. Upon actuation, the device discharges a measuredamount of therapeutic agent by compressed gas, thus affording a reliablemethod of administering a set amount of agent. DPI dispenses therapeuticagents in the form of a free flowing powder that can be dispersed in thepatient's inspiratory air-stream during breathing by the device. Inorder to achieve a free flowing powder, the therapeutic agent isformulated with an excipient such as lactose. A measured amount of thetherapeutic agent is stored in a capsule form and is dispensed with eachactuation.

Recently, pharmaceutical compositions have been developed for drugs thatshow poor bioavailability based upon the principle that bioavailabilitycan be increased by increasing the surface area i.e., decreasingparticle size. For example, U.S. Pat. No. 4,107,288 describes apharmaceutical formulation having particles in the size range from 10 to1,000 nm in which the active material is supported on a cross-linkedmatrix of macromolecules. U.S. Pat. No. 5,145,684 describes theproduction of a pharmaceutical formulation in which the drug substanceis pulverized to nanoparticles (average particle size of 400 nm) in thepresence of a surface modifier and then dispersed in a liquid medium togive a pharmaceutical formulation that exhibits remarkably highbioavailability.

The compositions are comprised of, in general, at least one chemicalentity described herein in combination with at least onepharmaceutically acceptable excipient. Acceptable excipients arenon-toxic, aid administration, and do not adversely affect thetherapeutic benefit of the at least one chemical entity describedherein. Such excipient may be any solid, liquid, semi-solid or, in thecase of an aerosol composition, gaseous excipient that is generallyavailable to one of skill in the art.

Solid pharmaceutical excipients include starch, cellulose, talc,glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silicagel, magnesium stearate, sodium stearate, glycerol monostearate, sodiumchloride, dried skim milk and the like. Liquid and semisolid excipientsmay be selected from glycerol, propylene glycol, water, ethanol andvarious oils, including those of petroleum, animal, vegetable orsynthetic origin, e.g., peanut oil, soybean oil, mineral oil, sesameoil, etc. Liquid carriers, for injectable solutions, include water,saline, aqueous dextrose, and glycols.

Compressed gases may be used to disperse a chemical entity describedherein in aerosol form. Inert gases suitable for this purpose arenitrogen, carbon dioxide, etc. Other suitable pharmaceutical excipientsand their formulations are described in Remington's PharmaceuticalSciences, edited by E. W. Martin (Mack Publishing Company, 18th ed.,1990).

The amount of the chemical entity in a composition can vary within thefull range employed by those skilled in the art. Typically, thecomposition will contain, on a weight percent (wt %) basis, from about0.01-99.99 wt % of at least one chemical entity described herein basedon the total composition, with the balance being one or more suitablepharmaceutical excipients. In certain embodiments, the at least onechemical entity described herein is present at a level of about 1-80 wt%.

In various embodiments, pharmaceutical compositions of the presentinvention encompass compounds of Formula (I), (Ia), (II) or (IIa), saltsthereof, and combinations of the above.

Synthetic Methods

The methods of synthesis may employ readily available starting materialsusing the following general methods and procedures. It will beappreciated that where typical or preferred process conditions (i.e.,reaction temperatures, times, mole ratios of reactants, solvents,pressures, etc.) are given; other process conditions can also be usedunless otherwise stated. Optimum reaction conditions may vary with theparticular reactants or solvent used, but such conditions can bedetermined by one skilled in the art by routine optimization procedures.

Additionally, the methods of this invention may employ protecting groupswhich prevent certain functional groups from undergoing undesiredreactions. Suitable protecting groups for various functional groups aswell as suitable conditions for protecting and deprotecting particularfunctional groups are well known in the art. For example, numerousprotecting groups are described in T. W. Greene and G. M. Wuts,Protecting Groups in Organic Synthesis, Third Edition, Wiley, New York,1999, and references cited therein.

Furthermore, the provided chemical entities may contain one or morechiral centers and such compounds can be prepared or isolated as purestereoisomers, i.e., as individual enantiomers or diastereomers, or asstereoisomer-enriched mixtures. All such stereoisomers (and enrichedmixtures) are included within the scope of this specification, unlessotherwise indicated. Pure stereoisomers (or enriched mixtures) may beprepared using, for example, optically active starting materials orstereoselective reagents well-known in the art. Alternatively, racemicmixtures of such compounds can be separated using, for example, chiralcolumn chromatography, chiral resolving agents and the like.

The starting materials for the following reactions are generally knowncompounds or can be prepared by known procedures or obviousmodifications thereof. For example, many of the starting materials areavailable from commercial suppliers such as Aldrich Chemical Co.(Milwaukee, Wis., USA), Bachem (Torrance, Calif., USA), Ernka-Chemce orSigma (St. Louis, Mo., USA). Others may be prepared by procedures, orobvious modifications thereof, described in standard reference textssuch as Fieser and Fieser's Reagents for Organic Synthesis, Volumes 1-15(John Wiley and Sons, 1991), Rodd's Chemistry of Carbon Compounds,Volumes 1-5 and Supplementals (Elsevier Science Publishers, 1989),Organic Reactions, Volumes 1-40 (John Wiley and Sons, 1991), March'sAdvanced Organic Chemistry, (John Wiley and Sons, 4th Edition), andLarock's Comprehensive Organic Transformations (VCH Publishers Inc.,1989).

Unless specified to the contrary, the reactions described herein maytake place at atmospheric pressure, generally within a temperature rangefrom −78° C. to 200° C. Further, except as employed in the Example or asotherwise specified, reaction times and conditions are intended to beapproximate, e.g., taking place at about atmospheric pressure within atemperature range of about −78° C. to about 110° C. over a period ofabout 1 to about 24 hours; reactions left to run overnight average aperiod of about 16 hours.

The terms “solvent,” “organic solvent,” and “inert solvent” each mean asolvent inert under the conditions of the reaction being described inconjunction therewith, including, for example, benzene, toluene,acetonitrile, tetrahydrofuranyl (“THF”), dimethylformamide (“DMF”),chloroform, methylene chloride (or dichloromethane), diethyl ether,methanol, N-methylpyrrolidone (“NMP”), pyridine and the like.

Isolation and purification of the chemical entities and intermediatesdescribed herein can be affected, if desired, by any suitable separationor purification procedure such as, for example, filtration, extraction,crystallization, column chromatography, thin-layer chromatography orthick-layer chromatography, or a combination of these procedures.Specific illustrations of suitable separation and isolation procedurescan be had by reference to the examples herein below. However, otherequivalent separation or isolation procedures can also be used.

When desired, the (R)- and (S)-isomers may be resolved by methods knownto those skilled in the art, for example by formation ofdiastereoisomeric salts or complexes which may be separated, forexample, by crystallization; via formation of diastereoisomericderivatives which may be separated, for example, by crystallization,gas-liquid or liquid chromatography; selective reaction of oneenantiomer with an enantiomer-specific reagent, for example enzymaticoxidation or reduction, followed by separation of the modified andunmodified enantiomers; or gas-liquid or liquid chromatography in achiral environment, for example on a chiral support, such as silica witha bound chiral ligand or in the presence of a chiral solvent.Alternatively, a specific enantiomer may be synthesized by asymmetricsynthesis using optically active reagents, substrates, catalysts orsolvents, or by converting one enantiomer to the other by asymmetrictransformation.

EXAMPLE AND GENERAL SYNTHESIS

The following example and prophetic synthesis method serve to more fullydescribe the manner of making and using the above-described invention.It is understood that this in no way serve to limit the true scope ofthe invention, but rather is presented for illustrative purposes. Unlessotherwise specified, the following abbreviations have the followingmeanings. If an abbreviation is not defined, it has its generallyaccepted meaning.

-   -   aq.=aqueous    -   μL=microliters    -   μM=micromolar    -   NMR=nuclear magnetic resonance    -   Boc=tert-butoxycarbonyl    -   br=broad    -   Cbz=benzyloxycarbonyl    -   d=doublet    -   ° C.═degrees Celsius    -   DCM=dichloromethane    -   dd=doublet of doublets    -   DMAP=N,N-dimethylaminopyridine    -   DMEM=Dulbeco's Modified Eagle's Medium    -   DMF=N,N-dimethylformamide    -   DMSO=dimethylsufoxide    -   EDC═N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride    -   EtOAc=ethyl acetate    -   g=gram    -   h or hr=hour(s)    -   HPLC═high performance liquid chromatography    -   Hz=Hertz    -   IU=International Units    -   IC₅₀=50% inhibitory concentration    -   J=coupling constant in Hz    -   LCMS=liquid chromatography mass spectrometry    -   m=multiplet    -   M=molar concentration    -   M+H⁺=parent mass spectrum peak plus H+    -   mg=milligram    -   min=minute(s)    -   mL=milliliter    -   mM=millimolar    -   mmol=millimole    -   MMTr=monomethoxytrityl    -   MS=mass spectrum    -   MTBE=methyl tert-butyl ether    -   nM=nanomolar    -   PE=petroleum ether    -   ppm=parts per million    -   q.s.=sufficient amount    -   s=singlet    -   RT=room temperature    -   sat.=saturated    -   t=triplet    -   TBDMS=tert-butyldimethylsilyl    -   TBDPS=tert-butyldiphenylsilyl    -   TEA=triethylamine    -   THF=tetrahydrofuran    -   TMS=trimethylsilyl

Additionally, various compounds of the invention may be made, in oneembodiment, by way of the general synthesis routes set forth in Schemes1-4 below:

wherein n is from 0 to 20, and each of R, R₁, R₂, R₃, R₄ and R₅ arealkyl.

Equipment Description

¹H NMR spectra were recorded on Varian or Bruker spectrometers. Chemicalshifts are expressed in parts per million (ppm, 6 units). Couplingconstants are in units of hertz (Hz). Splitting patterns describeapparent multiplicities and are designated as s (singlet), d (doublet),t (triplet), q (quartet), quint (quintet), m (multiplet), br (broad).

Representative equipment and conditions for acquiring analytical lowresolution LCMS are described below.

Instrumentation:

Waters Acquity UPLC-MS system with SQ detectors

MS Conditions:

Scan Mode: Alternating positive/negative electrospray

Scan Range: 125-1200 amu

Scan Time: 150 msecInterscan Delay: 50 msec

LC Conditions:

The UPLC analysis was conducted on a Phenomenex Kinetex 1.7 um2.1×50 mm XB-C18 column at 40° C.0.2 uL of sample was injected using PLNO (partial loop with needleoverfill) injection mode.The gradient employed was:

Mobile Phase A: Water+0.2% v/v Formic Acid Mobile Phase B:Acetonitrile+0.15% v/v Formic Acid

Time % A % B Flow Rate 0.00 min  95 5 1 ml/min 1.1 min 1 99 1 ml/min 1.5min 1 99 1 ml/minUV detection provided by summed absorbance signal from 210 to 350 nmscanning at 40 Hz.

Example 1:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylcyclohexanecarboxylate

Step A:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-2-ethynyltetrahydrofuran-3-ylacetate. A suspension of 2-fluoro-9H-purin-6-amine (0.545 g, 3.56 mmol)in anhydrous MeCN (10 mL) in a screw-capped glass pressure vessel undera nitrogen atmosphere was treated with trimethylsilyl2,2,2-trifluoro-N-(trimethylsilyl)acetimidate (1.89 ml, 7.12 mmol) andheated to 80° C. with stirring in an oil bath. After 45 minutes most ofthe solid had dissolved. The solution was treated with(4S,5R)-5-(((tert-butyldiphenylsilyl)oxy)methyl)-5-ethynyltetrahydrofuran-2,4-diyldiacetate (1.14 g, 2.37 mmol, prepared according to Org. Lett., Vol. 13,No. 19, 2011) dissolved in MeCN (9 mL) followed by freshly prepared 0.2Mtrifluoromethanesulfonic acid/MeCN (2.37 ml, 0.474 mmol) (prepared bydissolving 44 μL of triflic acid in 2.5 mL of MeCN). The temperature wasmaintained at 80° C. After 1.5 hour at 80° C. LCMS indicated completereaction. The solution was cooled to RT, quenched by addition of 1Maqueous HCl (3 mL). After stirring the mixture briefly, it waspartitioned between saturated aqueous NaHCO₃ and EtOAc and the phasesseparated. The aqueous phase was extracted with EtOAc (2×). The combinedEtOAc solutions were dried over Na₂SO₄ and concentrated at reducedpressure to give a tan solid. This material was subjected to flashchromatography (silica gel, 0-100% EtOAc/DCM) and the higher R_(f)component isolated to afford the title compound (0.63 g, 46%) as a whitesolid. LCMS (ESI) m/z calcd for C₃₀H₃₂FN₅O₄Si: 573.2. Found: 574.4(M+1)⁺. ¹H NMR (400 MHz, Methanol-d₄) δ 8.10 (s, 1H), 7.59-7.67 (m, 4H),7.26-7.45 (m, 6H), 6.39 (t, J=6.6 Hz, 1H), 5.91 (dd, J=7.0, 5.5 Hz, 1H),3.97 (d, J=10.9 Hz, 1H), 3.86 (d, J=10.9 Hz, 1H), 3.05-3.18 (m, 2H),2.64-2.74 (m, 1H), 2.14 (s, 3H), 0.97-1.04 (m, 9H).

Step B:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-2-ethynyltetrahydrofuran-3-ol.To a stirred solution of(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-2-ethynyltetrahydrofuran-3-ylacetate (0.62 g, 1.08 mmol) in 1:1 THF/MeOH (4 mL) was added 25%NaOMe/MeOH (3 drops). The resulting solution was stirred at RT. After 30minutes LCMS indicated complete reaction. The solution was treated withglacial AcOH (5 drops) and concentrated to dryness at reduced pressure.The residue was partitioned between 8:2 chloroform/iPrOH andhalf-saturated aqueous NaHCO₃ and the phases separated. The aqueousphase was extracted with two additional portions of 8:2chloroform/iPrOH. The combined organic solutions were dried over Na₂SO₄and concentrated to dryness at reduced pressure to afford the titlecompound (0.52 g, 91%) as a white solid. LCMS (ESI) m/z calcd forC₂₈H₃₀FN₅O₃Si: 531.2. Found: 532.3 (M+1)⁺. ¹H NMR (400 MHz, Methanol-d₄)δ 8.17 (s, 1H), 7.53-7.66 (m, 4H), 7.22-7.45 (m, 6H), 6.32 (dd, J=7.8,3.1 Hz, 1H), 5.01 (t, J=7.8 Hz, 1H), 3.87 (q, J=11.3 Hz, 2H), 3.05 (s,1H), 2.90-2.99 (m, 1H), 2.63-2.72 (m, 1H), 0.94 (s, 9H).

Step C:9-((2R,4S,5R)-5-(((tert-butyldiphenylsilyl)oxy)methyl)-5-ethynyl-4-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)-2-fluoro-N-((4-methoxyphenyl)diphenylmethyl)-9H-purin-6-amine.To a stirred suspension of(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-2-ethynyltetrahydrofuran-3-ol(0.510 g, 0.959 mmol) in DCM (8 mL) was added silver nitrate (0.489 g,2.88 mmol), 2,4,6-trimethylpyridine (0.766 ml, 5.76 mmol), and(chloro(4-methoxyphenyl)methylene)dibenzene (0.889 g, 2.88 mmol). Theresulting orange suspension was stirred at RT. After 2 hours LCMSindicated complete reaction. The mixture was diluted with EtOAc andfiltered through celite to remove solids. The filtrate was washed with10% aqueous citric acid (2×), saturated aqueous NaHCO₃ (2×), dried overNa₂SO₄ and concentrated at reduced pressure to give a pale yellow foam.This material was subjected to flash chromatography (silica gel, 0-100%EtOAc/hexanes) to afford the title compound (1.00 g, 97%) as a whitefoam. LCMS (ESI) m/z calcd for C₆₈H₆₂FN₅O₅Si: 1075.5. Found: 1076.7(M+1)⁺. ¹H NMR (400 MHz, CDCl₃) δ 7.12-7.62 (m, 35H), 6.98 (s, 1H),6.74-6.82 (m, 4H), 6.22 (t, J=6.6 Hz, 1H), 4.75 (t, J=5.9 Hz, 1H), 3.93(d, J=11.3 Hz, 1H), 3.86 (d, J=11.3 Hz, 1H), 3.77 (s, 3H), 3.75 (s, 3H),2.77 (s, 1H), 1.71 (t, J=6.3 Hz, 2H), 0.87 (s, 9H).

Step D:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methanol.To a stirred solution of9-((2R,4S,5R)-5-(((tert-butyldiphenylsilyl)oxy)methyl)-5-ethynyl-4-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)-2-fluoro-N-((4-methoxyphenyl)diphenylmethyl)-9H-purin-6-amine(0.99 g, 0.92 mmol) in THF (8 mL) was added 1 M TBAF/THF (1.38 ml, 1.38mmol) by dropwise addition. The resulting solution was stirred at RT.After 1 hour LCMS indicated complete reaction. The solution was treatedwith glacial AcOH (0.10 mL) and concentrated at reduced pressure. Theresidue was dissolved in MeOH/DCM and again concentrated to dryness. Theresidue was subjected to flash chromatography (silica gel, 0-100%EtOAc/hexanes) to afford the title compound (0.623 g, 81%) as a whitesolid. LCMS (ESI) m/z calcd for C₅₂H₄₄FN₅O₅: 837.3. Found: 838.6 (M+1)⁺.¹H NMR (400 MHz, DMSO-d₆) δ 8.04 (s, 1H), 8.00 (s, 1H), 7.47-7.54 m,4H), 7.12-7.38 (m, 20H), 6.79-6.88 (m, 4H), 6.04 (t, J=6.3 Hz, 1H), 5.15(t, J=6.1 Hz, 1H), 4.47 (t, J=6.1 Hz, 1H), 3.84 (s, 1H), 3.69 (s, 3H),3.67 (s, 3H), 3.49-3.57 (m, 1H), 3.38-3.47 (m, 1H), 1.63-1.72 (m, 1H),1.49-1.58 (m, 1H).

Step E:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methylcyclohexanecarboxylate. To a stirred solution of((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methanol(51 mg, 0.061 mmol), cyclohexanecarboxylic acid (15.60 mg, 0.122 mmol)and DMAP (7.44 mg, 0.061 mmol) in DCM (0.8 mL) was added EDC (35.0 mg,0.183 mmol), followed by DIPEA (0.053 mL, 0.304 mmol) at ambienttemperature. The mixture was allowed to stir overnight. The mixture wasconcentrated and then purified on silica gel (0-50% hexanes/EtOAc) toafford the title compound (53 mg, 90%) as a colorless residue. LCMS(ESI) m/z calcd for C₆₆H₆₈FN₅O₈Si: 1078. Found: 1079 (M+1)⁺. LCMS (ESI)m/z calcd for C₅₉H₅₄FN₅O₆: 947.4. Found: 948.8 (M+1)⁺

Step F:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylcyclohexanecarboxylate. To a stirred solution of((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methylcyclohexanecarboxylate (53 mg, 0.055 mmol) in DCM (0.8 mL) was addedformic acid (0.2 mL), followed by triethylsilane (0.018 mL, 0.11 mmol).The resulting orange solution was stirred for 60 minutes and was thenconcentrated. The residue was purified on silica gel (0-10% DCM/MeOH) toafford the title compound (17 mg, 78%) as a white solid. LCMS (ESI) m/zcalcd for C₁₉H₂₂FN₅O₄: 403.2. Found: 404.3 (M+1)⁺. ¹H NMR (400 MHz,DMSO-d₆) δ=8.25 (s, 1H), 7.81 (br s, 2H), 6.24 (dd, J=3.8, 8.1 Hz, 1H),5.76 (d, J=5.5 Hz, 1H), 4.75-4.69 (m, 1H), 4.41 (d, J=11.7 Hz, 1H), 4.06(d, J=11.9 Hz, 1H), 3.61 (s, 1H), 2.88-2.80 (m, 1H), 2.54-2.42 (m, 1H,overlapping DMSO peak), 2.20-2.10 (m, 1H), 1.80-1.69 (m, 1H), 1.66-1.45(m, 4H), 1.32-1.01 (m, 5H).

Example 2:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylbenzoate

The title compound was prepared according to example 1, substitutingbenzoic acid for cyclohexanecarboxylic acid in Step E. LCMS (ESI) m/zcalcd for C₁₉H₁₆FN₅O₄: 397.1. Found: 398.2 (M+1)⁺. ¹H NMR (400 MHz,DMSO-d₆) δ=8.27 (s, 1H), 7.91-7.86 (m, 2H), 7.81 (br s, 2H), 7.68-7.62(m, 1H), 7.52-7.44 (m, 2H), 6.29 (dd, J=3.9, 8.2 Hz, 1H), 5.84 (d, J=5.5Hz, 1H), 4.92-4.84 (m, 1H), 4.61 (d, J=11.9 Hz, 1H), 4.40 (d, J=11.7 Hz,1H), 3.67 (s, 1H), 2.91-2.83 (m, 1H), 2.57-2.47 (m, 1H, overlapping DMSOpeak).

Example 3:[(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolan-2-yl]methyl2-phenylacetate

The title compound was prepared according to example 1, substituting2-phenylacetic acid for cyclohexanecarboxylic acid in Step E. LCMS (ESI)m/z calcd for C₂₀H₁₈FN₅O₄: 411.1. Found: 412.6 (M+1)⁺. ¹H NMR (400 MHz,DMSO-d₆) δ=8.27 (s, 1H), 7.83 (br s, 2H), 7.29-7.20 (m, 3H), 7.19-7.13(m, 2H), 6.26 (dd, J=4.1, 8.1 Hz, 1H), 5.77 (d, J=5.5 Hz, 1H), 4.76-4.68(m, 1H), 4.44 (d, J=11.7 Hz, 1H), 4.15 (d, J=11.7 Hz, 1H), 3.71-3.48 (m,3H), 2.83-2.74 (m, 1H), 2.56-2.43 (m, 1H, overlapping DMSO peak).

Example 4:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl2-phenylacetate

The title compound was prepared according to example 1, substitutingtrans-4-(tert-butyl)cyclohexanecarboxylic acid for cyclohexanecarboxylicacid in Step E. LCMS (ESI) m/z calcd for C₂₃H₃₀FN₅O₄: 459.2. Found:460.3 (M+1)⁺. ¹H NMR (400 MHz, DMSO-d₆) δ=8.25 (s, 1H), 7.80 (br s, 2H),6.23 (dd, J=3.9, 8.0 Hz, 1H), 5.75 (d, J=5.5, 1H), 4.76-4.69 (m, 1H),4.42 (d, J=11.7 Hz, 1H), 4.05 (d, J=11.7 Hz, 1H), 3.61 (s, 1H),2.88-2.80 (m, 1H), 2.54-2.42 (m, 1H, overlapping DMSO peak), 2.10-1.96(m, 1H), 1.91-1.80 (m, 1H), 1.75-1.59 (m, 3H), 1.26-1.03 (m, 2H),0.98-0.84 (m, 3H), 0.81 (s, 9H).

Example 5:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyltrans-4-pentylcyclohexane-1-carboxylate

The title compound was prepared according to example 1, substitutingtrans-4-pentylcyclohexanecarboxylic acid for cyclohexanecarboxylic acidin Step E. LCMS (ESI) m/z calcd for C₂₄H₃₃FN₅O₄: 473.2. Found: 474.4(M+1)⁺. ¹H NMR (400 MHz, DMSO-d₆) δ=8.25 (s, 1H), 7.81 (br s, 2H), 6.23(dd, J=3.9, 8.0 Hz, 1H), 5.75 (d, J=5.5 Hz, 1H), 4.76-4.68 (m, 1H), 4.42(d, J=11.9 Hz, 1H), 4.05 (d, J=11.7 Hz, 1H), 3.61 (s, 1H), 2.87-2.79 (m,1H), 2.54-2.42 (m, 1H, overlapping DMSO peak), 2.11-2.02 (m, 1H),1.85-1.75 (m, 1H), 1.73-1.57 (m, 3H), 1.35-1.04 (m, 11H), 0.92-0.74 (m,5H).

Example 6:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl2-(2-methoxyethoxy)acetate

The title compound was prepared according to example 1, substituting2-(2-methoxyethoxy)acetic acid for cyclohexanecarboxylic acid in Step E.LCMS (ESI) m/z calcd for C₁₇H₂₀FN₅O₆: 409.1. Found: 410.3 (M+1)⁺. ¹H NMR(400 MHz, DMSO-d₆) δ=8.27 (s, 1H), 7.83 (br s, 2H), 6.25 (dd, J=4.3, 7.9Hz, 1H), 5.78 (d, J=5.5 Hz, 1H), 4.72-4.65 (m, 1H), 4.48 (d, J=11.7 Hz,1H), 4.18 (d, J=11.7 Hz, 1H), 4.12 (d, J=16.7 Hz, 1H), 3.99 (d, J=16.7Hz, 1H), 3.64 (s, 1H), 3.54-3.48 (m, 2H), 3.44-3.38 (m, 2H), 3.21 (s,3H), 2.82-2.74 (m, 1H), 2.54-2.43 (m, 1H, overlapping DMSO peak).

Example 7:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl2-(2-(2-methoxyethoxy)ethoxy)acetate

The title compound was prepared according to example 1, substituting2-(2-(2-methoxyethoxy)ethoxy)acetic acid for cyclohexanecarboxylic acidin Step E. LCMS (ESI) m/z calcd for C₁₉H₂₄FN₅O₇: 453.2. Found: 454.7(M+1)⁺. ¹H NMR (400 MHz, DMSO-d₆) δ=8.27 (s, 1H), 7.82 (br s, 2H), 6.25(dd, J=4.1, 7.9 Hz, 1H), 5.78 (d, J=5.5 Hz, 1H), 4.71-4.64 (m, 1H), 4.48(d, J=11.7 Hz, 1H), 4.18 (d, J=11.7 Hz, 1H), 4.12 (d, J=16.9 Hz, 1H),4.00 (d, J=16.7 Hz, 1H), 3.63 (s, 1H), 3.54-3.45 (m, 6H), 3.43-3.36 (m,2H), 3.21 (s, 3H), 2.81-2.73 (m, 1H), 2.53-2.43 (m, 1H, overlapping DMSOpeak).

Example 8:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl2-(2-butoxyethoxy)acetate

The title compound was prepared according to example 1, substituting2-(2-butoxyethoxy)acetic acid for cyclohexanecarboxylic acid in Step E.LCMS (ESI) m/z calcd for C₂₀H₂₆FN₅O₆: 451.2. Found: 452.3 (M+1)⁺. ¹H NMR(400 MHz, DMSO-d₆) δ=8.28 (s, 1H), 7.84 (br s, 2H), 6.26 (dd, J=4.2, 7.7Hz, 1H), 5.79 (d, J=5.5 Hz, 1H), 4.72-4.65 (m, 1H), 4.49 (d, J=11.7 Hz,1H), 4.19 (d, J=11.7 Hz, 1H), 4.14 (d, J=16.7 Hz, 1H), 4.02 (d, J=16.7Hz, 1H), 3.64 (s, 1H), 3.55-3.49 (m, 2H), 3.48-3.43 (m, 2H), 3.37-3.32(m, 2H), 2.82-2.74 (m, 1H), 2.54-2.44 (m, 1H, overlapping DMSO peak),1.49-1.37 (m, 2H), 1.35-1.20 (m, 2H), 0.90-0.80 (m, 3H).

Example 9:10-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)-10-oxodecanoicacid

Step A:1-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl)10-tert-butyl) decanedioate. 10-(tert-butoxy)-10-oxodecanoic acid (96mg, 0.37 mmol) was dissolved in DMF (1 mL), EDC (286 mg, 1.50 mmol) andDMAP (182 mg, 1.489 mmol) were added, the resulting mixture was stirredfor 2 h at RT. Then,((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methanol(260 mg, 0.310 mmol) was addded, and the resulting mixture was stirredfor 1 h at RT. The reaction mixture was added mixed water (10 mL) andextracted with EtOAc (3×5 mL). The combined organic phases were washedwith brine (10 mL), dried over Na₂SO₄, and concentrated under vacuum.The residue was purified by preparative TLC (MeOH:DCM=1:20) to affordthe title compound (89 mg, 26%) as a white solid. LCMS (ESI) m/z calcdfor C₂₆H₃₆FN₅O₆: 533. Found: 534 (M+1)⁺.

Step B:10-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)-10-oxodecanoicacid. 1-(tert-butyl) 10-(((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methyl)decanedioate (115 mg, 0.107 mmol) was dissolved in DCM (3 mL). TFA(0.158 mL, 2.13 mmol) was added, and the resulting mixture was stirredat RT. After 1 h the solution was concentrated to dryness at reducedpressure and the residue purified by RP-HPLC (C₁₈, MeCN/water with 0.1%formic acid) to afford the title compound (15.6 mg, 31%) as a whitesolid. LCMS (ESI) m/z calcd for C₂₂H₂₈FN₅O₆: 477. Found: 478 (M+1)⁺. ¹HNMR (400 MHz, DMSO-d₆) δ 10.90 (s, 1H), 8.27 (s, 1H), 7.86 (d, J=6.0 Hz,2H), 6.23 (dd, J=8.0, 4.4 Hz, 1H), 4.71 (t, J=5.2 Hz, 1H), 4.42 (d,J=11.6 Hz, 1H), 4.08 (d, J=11.6 Hz, 1H), 3.63 (s, 1H), 2.80-2.77 (m,1H), 2.47 (dd, J=8.4, 3.0 Hz, 1H), 2.20-2.14 (m, 4H), 1.48-1.39 (m, 4H),1.18 (d, J=9.4 Hz, 8H).

Example 10:14-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)-14-oxotetradecanoicacid

The title compound was prepared according to example 9, substituting14-(tert-butoxy)-14-oxotetradecanoic acid for10-(tert-butoxy)-10-oxodecanoic acid in step A. LCMS (ESI) m/z calcd forC₂₆H₃₆FN₅O₆: 533. Found: 534 (M+1)⁺. ¹H NMR (400 MHz, Methanol-d₄) δ8.15 (s, 1H), 6.30 (dd, J=3.6, 8.0 Hz, 1H), 4.87 (s, 1H), 4.47 (d,J=12.0 Hz, 1H), 4.25 (d, J=12.0 Hz, 1H), 3.17 (s, 1H), 2.91-2.88 (m,1H), 2.70-2.64 (m, 1H), 2.29-2.18 (m, 4H), 1.63-1.55 (m, 2H), 1.51-1.43(m, 2H), 1.26 (m, 16H).

Example 11:20-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)-20-oxoicosanoicacid

The title compound was prepared according to example 9, substituting20-(tert-butoxy)-20-oxoicosanoic acid for10-(tert-butoxy)-10-oxodecanoic acid in step A. LCMS (ESI) m/z calcd forC₃₂H₄₈FN₅O₆: 617. Found: 618 (M+1)⁺. ¹H NMR (400 MHz, Methanol-d₄) δ8.15 (s, 1H), 6.30 (dd, J=3.6, 8.0 Hz, 1H), 4.87 (s, 1H), 4.47 (d,J=12.0 Hz, 1H), 4.25 (d, J=12.0 Hz, 1H), 3.17 (s, 1H), 2.93-2.88 (m,1H), 2.70-2.62 (m, 1H), 2.29-2.15 (m, 4H), 1.63-1.55 (m, 2H), 1.52-1.44(m, 2H), 1.28 (m, 28H).

Example 12:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl(1,3-bis(heptanoyloxy)propan-2-yl) glutarate

Step A: 2-oxopropane-1,3-diyl diheptanoate. To a suspension of1,3-dihydroxypropan-2-one (500 mg, 5.55 mmol) in DCM (20 mL) was addedpyridine (0.940 mL, 11.6 mmol) and heptanoyl chloride (1.76 mL, 11.4mmol) and the mixture was stirred at ambient temperature for 18 h. Themixture was diluted with DCM and washed with water. The organic phasewas washed with brine, dried (Na₂SO₄), concentrated and purified onsilica gel (EtOAc/hexanes 0-20%) to provide 2-oxopropane-1,3-diyldiheptanoate (1.17 g, 66%) as an off-white solid. LCMS (ESI) m/z calcdfor C₁₇H₃₂O₅: 316. Found: 315 (M−1)⁺. ¹H NMR (400 MHz, Chloroform-d): δ4.77 (s, 4H), 2.45 (t, J=7.5 Hz, 4H), 1.61-1.77 (m, 4H), 1.23-1.43 (m,12H), 0.85-0.99 (m, 6H).

Step B: 2-hydroxypropane-1,3-diyl diheptanoate. To a solution of2-oxopropane-1,3-diyl diheptanoate (500 mg, 1.59 mmol) in THF (10 mL)was added water (0.5 mL and the mixture was cooled to 0° C. then NaBH₄(36.1 mg, 0.954 mmol) was added and stirring under nitrogen atmospherecontinued for 1 h and then 1 h at ambient temperature. SaturatedNH₄Cl/water was added and the mixture was extracted with EtOAc. Theorganic phase was dried (Na₂SO₄), concentrated and purified on silicagel (EtOAc/hexanes 0-100%) to provide 2-hydroxypropane-1,3-diyldiheptanoate (487 mg, 64%) as a clear oil. ¹H NMR (400 MHz,Chloroform-d): δ 4.03-4.29 (m, 5H), 2.42-2.51 (m, 1H), 2.37 (t, J=7.5Hz, 4H), 1.61-1.74 (m, 4H), 1.24-1.44 (m, 12H), 0.86-0.98 (m, 6H).

Step C: 5-((1,3-bis(heptanoyloxy)propan-2-yl)oxy)-5-oxopentanoic acid.To a solution of 2-hydroxypropane-1,3-diyl diheptanoate (160 mg, 0.506mmol) in DCM (1 mL)/tetrahydrofuran (1 mL)/pyridine (1 mL) was addedDMAP (6.18 mg, 0.051 mmol) followed by dihydro-2H-pyran-2,6(3H)-dione(115 mg, 1.01 mmol) and the mixture was heated to 60° C. for 6.5 h. Themixture was diluted with EtOAc and washed with 1 M HCl/water. Theorganic phase was dried (Na₂SO₄), concentrated and purified on silicagel (MeOH/dichloromethane 0-5%) to provide5-((1,3-bis(heptanoyloxy)propan-2-yl)oxy)-5-oxopentanoic acid (190 mg,87%) as a clear oil. LCMS (ESI) m/z calcd for C₂₂H₃₃O₈: 430. Found: 431(M−1)⁺. ¹H NMR (400 MHz, Chloroform-d): δ 5.21-5.35 (m, 1H), 4.28-4.42(m, 2H), 4.09-4.25 (m, 2H), 2.40-2.53 (m, 4H), 2.27-2.38 (m, 4H), 1.99(t, J=7.3 Hz, 2H), 1.51-1.73 (m, 4H), 1.25-1.43 (m, 12H), 0.85-0.98 (m,6H).

Step D:9-((2R,4S,5R)-4-((tert-butyldimethylsilyl)oxy)-5-(((tert-butyldimethylsilyl)oxy)methyl)-5-ethynyltetrahydrofuran-2-yl)-2-fluoro-9H-purin-6-amine.To a suspension of(2R,3S)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (268 mg, 0.914 mmol) (Org. Lett., Vol. 13, No. 19,2011, 5264-5299), in DMF (2 mL) was added imidazole (311 mg, 4.57 mmol)and TBDMS-Cl (344 mg, 2.285 mmol) and the mixture was stirred at ambienttemperature for 1 h and then at 50° C. for 1 h. LCMS showed mono-silylproduct only so more imidazole (311 mg, 4.57 mmol) and TBDMS-Cl (344 mg,2.285 mmol) was added and the mixture was stirred at 50° C. undernitrogen atmosphere for 1 h. Water was added and the mixture was stirredat ambient temperature for 18 h. The mixture was filtered and the stickysolid residue was dissolved in DCM, dried (Na₂SO₄), concentrated andpurified on silica gel (EtOAc/hexanes 20-100%) to provide9-((4S,5R)-4-((tert-butyldimethylsilyl)oxy)-5-(((tert-butyldimethylsilyl)oxy)methyl)-5-ethynyltetrahydrofuran-2-yl)-2-fluoro-9H-purin-6-amine (430mg, 90%) as an off-white solid. LCMS (ESI) m/z calcd for C₂₄H₄₀FN₅O₃Si₂:521. Found: 522 (M+1)⁺. ¹H NMR (400 MHz, Chloroform-d): δ 8.01-8.10 (m,1H), 6.38 (dd, J=7.2, 4.3 Hz, 1H), 5.80 (br s, 2H), 4.84 (t, J=7.0 Hz,1H), 3.98 (d, J=11.2 Hz, 1H), 3.82 (d, J=11.2 Hz, 1H), 2.59-2.79 (m,2H), 2.56 (s, 1H), 0.96 (s, 9H), 0.91 (s, 9H), 0.16 (s, 3H), 0.15 (s,3H), 0.10 (s, 3H), 0.06 (s, 3H).

Step E:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methanol.To a solution of9-((4S,5R)-4-((tert-butyldimethylsilyl)oxy)-5-(((tert-butyldimethylsilyl)oxy)methyl)-5-ethynyltetrahydrofuran-2-yl)-2-fluoro-9H-purin-6-amine(32 mg, 0.061 mmol) in MeOH (0.7 mL) was added phosphomolybdic acidhydrate (56.0 mg, 6.13 μmol) and the mixture was stored at −5° C. for 18h. The mixture was filtered and the yellowish solid washed with MeOH andDCM. The filtrate was concentrated and purified on silica gel(EtOAc/hexanes 20-100%) to provide((2R,3S)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methanol(16.4 mg, 65%) as an off-white solid. LCMS (ESI) m/z calcd forC₁₈H₂₆FN₅O₃Si: 407. Found: 408 (M+1)⁺. ¹H NMR (400 MHz, DMSO-d₆): δ7.74-8.04 (m, 2H), 6.26 (dd, J=6.9, 5.7 Hz, 1H), 5.30 (dd, J=6.7, 5.5Hz, 1H), 4.77 (t, J=6.2 Hz, 1H), 3.62-3.73 (m, 1H), 3.47-3.57 (m, 2H),2.84 (dt, J=12.5, 6.4 Hz, 1H), 2.50-2.52 (m, 1H), 2.35-2.45 (m, 1H),0.89-0.97 (m, 9H), 0.14 (d, J=3.3 Hz, 6H).

Step F:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methyl(1,3-bis(heptanoyloxy)propan-2-yl) glutarate. To a suspension of((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methanol (11.7 mg, 0.029 mmol) in DCM(1.5 mL) was added5-((1,3-bis(heptanoyloxy)propan-2-yl)oxy)-5-oxopentanoic acid (25 mg,0.057 mmol) followed by DMAP (3.5 mg, 0.029 mmol), EDC (16.5 mg, 0.0860mmol) and DIEA (0.025 mL, 0.144 mmol) and the cloudy mixture was stirredat ambient temperature for 3.5 h. The mixture was diluted with DCM andwashed with water. The organic phase was dried (Na₂SO₄), concentratedand purified on silica gel (EtOAc/hexanes 0-40-100%) to provide((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methyl(1,3-bis(heptanoyloxy)propan-2-yl) glutarate (17.8 mg, 76%). LCMS (ESI)m/z calcd for C₄₀H₆₂FN₅O₁₀Si: 819. Found: 820 (M+1)⁺. ¹H NMR (400 MHz,Chloroform-d): δ 7.86 (s, 1H), 6.23-6.30 (m, 1H), 5.87-6.03 (m, 2H),5.28 (ddd, J=5.7, 4.4, 1.3 Hz, 1H), 4.97 (t, J=7.51 Hz, 1H), 4.50 (d,J=12.2 Hz, 1H), 4.33 (dt, J=11.9, 4.4 Hz, 2H), 4.26 (d, J=11.9 Hz, 1H),4.18 (ddd, J=11.9, 5.90, 1.6 Hz, 2H), 2.85-2.97 (m, 1H), 2.63 (s, 2H),2.28-2.44 (m, 8H), 1.84-1.98 (m, 2H), 1.57-1.68 (m, 4H), 1.25-1.39 (m,12H), 0.94-0.99 (m, 9H), 0.86-0.93 (m, 6H), 0.17 (s, 3H), 0.16 (s, 3H).

Step G:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl(1,3-bis(heptanoyloxy)propan-2-yl) glutarate. To a solution of((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methyl(1,3-bis(heptanoyloxy)propan-2-yl) glutarate (17 mg, 0.021 mmol) intetrahydrofuran (0.7 mL) at 0° C. was added ˜ 50 uL of a solutionprepared by adding acetic acid (58 uL, 1 mmol) to 1M TBAF/THF (1 mL, 1mmol). The mixture was stored at 0° C. for 18 h then stirred 2 h atambient temperature. The mixture was concentrated and purified on silicagel (EtOAc/hexanes 0-100%) to provide((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl(1,3-bis(heptanoyloxy)propan-2-yl) glutarate (10.3 mg, 70% yield) as asticky solid. LCMS (ESI) m/z calcd for C₃₄H₄₈FN₅O₁₀: 705. Found: 706(M+1)⁺. ¹H NMR (400 MHz, Chloroform-d): δ 7.84-7.94 (m, 1H), 6.32 (dd,J=7.5, 4.4 Hz, 1H), 5.85 (br s, 2H), 5.21-5.36 (m, 1H), 4.85 (q, J=7.07Hz, 1H), 4.47 (s, 2H), 4.35 (ddd, J=11.9, 4.4, 2.0 Hz, 2H), 4.18 (ddd,J=11.9, 5.7, 3.3 Hz, 2H), 2.94-3.12 (m, 1H), 2.81 (s, 1H), 2.68 (dd,J=7.4, 6.2 Hz, 2H), 2.39-2.54 (m, 4H), 2.34 (t, J=7.51 Hz, 4H),1.87-2.08 (m, 2H), 1.50-1.67 (m, 4H), 1.23-1.42 (m, 12H), 0.83-0.96 (m,6H).

Example 13:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl(1,3-bis(tetradecanoyloxy)propan-2-yl) glutarate

The title compound was prepared according to example 12, substitutingtetradecanoyl chloride for heptanoyl chloride in step A. LCMS (ESI) m/zcalcd for C₄₈H₇₆FN₅O₁₀: 902. Found: 903 (M+1)⁺. ¹H NMR (400 MHz,Chloroform-d): δ 7.89 (s, 1H), 6.32 (dd, J=7.5, 4.4 Hz, 1H), 5.95 (d,J=1.0 Hz, 2H), 5.19-5.38 (m, 1H), 4.84 (br s, 1H), 4.47 (s, 2H), 4.34(ddd, J=11.9, 4.5, 2.3 Hz, 2H), 4.18 (ddd, J=11.9, 5.7, 3.3 Hz, 2H),2.92-3.10 (m, 1H), 2.81 (s, 1H), 2.68 (d, J=13.6 Hz, 2H), 2.38-2.56 (m,4H), 2.34 (t, J=7.5 Hz, 4H), 1.88-2.08 (m, 2H), 1.59-1.64 (m, 4H),1.19-1.40 (m, 40H), 0.83-0.97 (m, 6H).

Example 14:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyltetradecyl glutarate

The title compound was obtained as a by-product from example 13, step GLCMS (ESI) m/z calcd for C₃₁H₄₆FN₅O₆: 603. Found: 604 (M+1)⁺. ¹H NMR(400 MHz, Methanol-d₄): δ 8.16 (s, 1H), 6.33 (dd, J=7.9, 3.8 Hz, 1H),4.87 (t, J=7.9 Hz, 1H), 4.50 (d, J=11.9 Hz, 1H), 4.31 (d, J=11.7 Hz,1H), 4.06 (t, J=6.7 Hz, 2H), 3.17 (s, 1H), 2.92 (ddd, J=13.6, 7.3, 3.7Hz, 1H), 2.68 (dt, J=13.7, 8.2 Hz, 1H), 2.23-2.47 (m, 4H), 1.84 (td,J=7.3, 1.3 Hz, 2H), 1.52-1.67 (m, 2H), 1.30 (s, 22H), 0.82-1.00 (m, 3H).

Example 15:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((ethoxycarbonyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methyl decanoate

Step A:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-2-ethynyltetrahydrofuran-3-ylethyl carbonate. To a stirred solution of(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-2-ethynyltetrahydrofuran-3-ol(51 mg, 0.096 mmol), triethylamine (0.067 mL, 0.480 mmol) and DMAP(11.72 mg, 0.096 mmol) in DCM (0.8 mL) at 0° C. was added ethylcarbonochloridate (0.018 mL, 0.192 mmol) in DCM (82 uL). The mixture wasstirred at 0° C. for 5 minutes, then stirred at ambient temperature for2 hours. LCMS of the reaction mixture indicated a mixture of tri-, di-and monosubstituted compounds. The mixture was diluted with EtOAc,washed with sat'd NaHCO₃ mixed with brine, followed by brine only andfinally dried (Na₂SO₄), filtered and concentrated. The residue waspurified on silica gel (0-50% DCM/EtOAc) to afford the title compound(15 mg, 26%) as a colorless residue. LCMS (ESI) m/z calcd forC₃₁H₃₄FN₅O₅Si: 603.2. Found: 604.4 (M+1)⁺.

Step B:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ylethyl carbonate. To a stirred solution of(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-2-ethynyltetrahydrofuran-3-ylethyl carbonate (15 mg, 0.025 mmol) in THF (0.5 mL) at ambienttemperature was added TBAF, 1M solution in THF (0.033 mL, 0.033 mmol)and the mixture was allowed to stir for 15 minutes. AcOH (5 drops) wasadded, stirred for several minutes and the mixture was thenconcentrated. The residue was diluted with EtOAc, washed with sat'dNaHCO₃, then brine, dried over Na₂SO₄, filtered and concentrated to acolorless residue which was used crude in the next step. LCMS (ESI) m/zcalcd for C₁₅H₁₆FN₅O₅: 365.1. Found: 366.2 (M+1)⁺.

Step C:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((ethoxycarbonyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methyldecanoate. To a solution of the crude product from the previous step,decanoic acid (8.61 mg, 0.050 mmol) and DMAP (3.05 mg, 0.025 mmol) inDCM (0.5 mL) at ambient temperature was added EDC (14.38 mg, 0.075mmol), followed by DIPEA (0.022 mL, 0.125 mmol) and the mixture wasallowed to stir overnight. The mixture was concentrated and thenpurified by RP-HPLC purification (C18, 10-100% MeCN/water with 0.1% FA)to afford the title compound (4.5 mg, 34% over 2 steps) as a whitesolid. LCMS (ESI) m/z calcd for C₂₅H₃₄FN₅O₆: 519.3. Found: 520.3 (M+1)⁺.¹H NMR (400 MHz, DMSO-d₆) δ 8.32 (s, 1H), 8.07-7.76 (m, 2H), 6.37-6.31(m, 1H), 5.57 (dd, J=5.3, 6.8 Hz, 1H), 4.42 (d, J=11.7 Hz, 1H),4.28-4.13 (m, 3H), 3.83 (s, 1H), 3.24-3.15 (m, 1H), 2.73-2.61 (m, 1H),2.35-2.14 (m, 2H), 1.50-1.38 (m, 2H), 1.31-1.11 (m, 15H), 0.84 (t, J=6.8Hz, 3H).

Example 16:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((heptanoyloxy)methyl)tetrahydrofuran-3-ylheptanoate

To a stirred solution of(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol(54 mg, 0.184 mmol) (Org. Lett., Vol. 13, No. 19, 2011, 5264-5299),heptanoic acid (0.104 mL, 0.737 mmol) and DMAP (45.0 mg, 0.368 mmol) inDMF (1.6 mL) at ambient temperature was added EDC (176 mg, 0.921 mmol)followed by DIPEA (0.322 mL, 1.841 mmol) and the mixture was allowed tostir overnight. Water was added and the mixture was extracted withEtOAc. The combined extracts were washed with water, then brine, driedover Na₂SO₄, filtered and concentrated. The residue was purified onsilica gel (0-10% DCM/MeOH) to afford the title compound (80 mg, 84%) asa white solid. LCMS (ESI) m/z calcd for C₂₆H₃₆FN₅O₅: 517.3. Found: 518.4(M+1)⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 8.33 (s, 1H), 8.05-7.75 (m, 2H),6.37-6.31 (m, 1H), 5.73-5.67 (m, 1H), 4.40 (d, J=11.7 Hz, 1H), 4.22 (d,J=11.7 Hz, 1H), 3.79 (s, 1H), 3.19-3.10 (m, 1H), 2.66-2.56 (m, 1H),2.43-2.15 (m, 4H), 1.63-1.53 (m, 2H), 1.49-1.38 (m, 2H), 1.36-1.10 (m,12H), 0.91-0.79 (m, 6H).

Example 17:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((tetradecanoyloxy)methyl)tetrahydrofuran-3-yltetradecanoate

The title compound was prepared according to example 16, substitutingtetradecanoic acid for heptanoic acid. LCMS (ESI) m/z calcd forC₄₀H₆₄FN₅O₅: 713.5. Found: 714.5 (M+1)⁺. ¹H NMR (400 MHz, Chloroform-d)δ 7.96 (s, 1H), 6.39 (t, J=6.4 Hz, 1H), 6.16 (br s, 2H), 5.61-5.66 (m,1H), 4.48 (d, J=12.1 Hz, 1H), 4.38 (J=12.1 Hz, 1H), 2.98 (dt, J=13.8,6.6 Hz, 1H), 2.64-2.74 (m, 2H), 2.27-2.46 (m, 4H), 1.55-1.69 (m, 4H),1.20-1.32 (m, 40H), 0.86 (t, J=6.64 Hz, 6H).

Example 18:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((decanoyloxy)methyl)-2-ethynyltetrahydrofuran-3-yldecanoate

Synthesis A

The title compound was prepared according to example 16, substitutingdecanoic acid for heptanoic acid. LCMS (ESI) m/z calcd for C₃₂H₄₈FN₅O₅:601.4. Found: 602.7 (M+1)⁺. ¹H NMR (400 MHz, Chloroform-d) δ 7.96 (s,1H), 6.39 (t, J=6.4 Hz, 1H), 6.14-6.33 (m, 2H), 5.64 (dd, J=7.4, 5.5 Hz,1H), 4.48 (d, J=12.1 Hz, 1H), 4.38 (d, J=12.1 Hz, 1H), 2.98 (dt, J=13.5,6.9 Hz, 1H), 2.62-2.74 (m, 2H), 2.27-2.42 (m, 4H), 1.55-1.69 (m, 4H),1.17-1.35 (m, 24H), 0.80-0.90 (m, 6H).

Synthesis B: Large scale preparation: To a solution of(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol(4.75 g, 16.2 mmol), TEA (9.03 mL, 64.8 mmol) and DMAP (0.396 g, 3.24mmol) in DCM (100 mL) was added decanoyl chloride (6.80 g, 35.6 mmol)dropwise. The resulting mixture was stirred for 2 hours at roomtemperature. LCMS indicated completion of reaction. The reaction wasquenched with water (100 ml) and extracted with DCM (100 ml×2). Thecombined organic phases were dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure. The reaction was repeated for fourtimes and the combined residues were purified by flash chromatography(silica gel, 660 g, pet. ether—0-80% EtOAc) to give the desired product(purity: 95%) as a white solid. The solid was triturated with EtOAc (300ml), stirred for 0.5 h, filtered through a Buchner funnel, rinsed withEtOAc, and dried under sun lamp for 6 h (T=50° C.). This solid wasstored in a cool dry place overnight to give crystalline(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((decanoyloxy)methyl)-2-ethynyltetrahydrofuran-3-yldecanoate (34 g, 99.43%, yield: 87%) as a white solid. LCMS (ESI) m/zcalcd for C₃₂H₄₈FN₅O₅: 601. Found: 602 (M+1)⁺. ¹H NMR (400 MHz,Chloroform-d) δ 7.93 (s, 1H), 6.40 (t, J=6.4 Hz, 1H), 6.13 (br s, 2H),5.65 (dd, J=6.8, 5.6 Hz, 1H), 4.49 (d, J=12.0 Hz, 1H), 4.39 (d, J=12.0Hz, 1H), 3.02-3.00 (m, 1H), 2.78-2.67 (m, 1H), 2.66 (s, 1H), 2.43-2.30(m, 4H), 1.67-1.58 (m, 4H), 1.40-1.17 (m, 24H), 0.89-0.86 (m, 6H).

Example 19:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((pentanoyloxy)methyl)tetrahydrofuran-3-yltridecanoate

Step A:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methylpentanoate. To a stirred solution of((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxy-phenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methanol (1.50 g, 1.79 mmol) and DMAP (0.109 g,0.895 mmol) in DCM (15 mL) at 0° C. was added TEA (0.749 mL, 5.37 mmol)followed by a solution of pentanoyl chloride (0.234 mL, 1.97 mmol) inDCM (1 mL). The resulting solution was stirred at 0° C. for 10 minutesand at RT for 30 minutes. LCMS indicated complete reaction. The reactionmixture was quenched with water, extracted with EtOAc (3×20 mL), theorganic phase was combined, washed with brine, dried over Na₂SO₄ andconcentrated under vacuum. The residue was subjected to preparative—TLC(MeOH:DCM=1:20) to give the desired product (1.5 g, 88%) as a whitesolid. LCMS (ESI) m/z calcd for C₅₇H₅₂FN₅O₆: 921. Found: 922 (M+1)⁺. ¹HNMR (400 MHz, DMSO-d₆) δ 8.12-7.91 (m, 2H), 7.51 (d, J=7.6 Hz, 4H),7.42-7.11 (m, 20H), 6.84 (d, J=8 Hz, 4H), 6.13 (s, 1H), 4.68 (s, 1H),4.06-3.95 (m, 2H), 3.85-3.77 (m, 1H), 3.74-3.54 (m, 6H), 2.10-1.99 (m,2H), 1.95-1.87 (m, 2H), 1.24-1.18 (m, 2H), 1.10-1.01 (m, 2H), 0.78-0.67(m, 3H).

Step B:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylpentanoate.((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methylpentanoate (1.46 g, 1.58 mmol) was dissolved in DCM (14 mL) and TFA (3mL), the resulting mixture was stirred for 1 h at RT. LCMS indicatedcomplete reaction. The reaction mixture was diluted with MeOH (14 mL)and concentrated under vacuum. The residue was subjected to reversephase HPLC purification (C18, 20-60% ACN/water with 0.1% FA) to give thedesired product as a white solid (525 mg, 87%). LCMS (ESI) m/z calcd forC₁₇H₂₀FN₅O₄: 377. Found: 378 (M+1)⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 8.28(s, 1H), 7.87 (br, 2H), 6.24 (dd, J=8.0, 3.9 Hz, 1H), 5.80 (d, J=5.4 Hz,1H), 4.74-4.69 (m, 1H), 4.43 (d, J=11.6 Hz, 1H), 4.09 (d, J=12.0 Hz,1H), 3.64 (s, 1H), 2.83-2.78 (m, 1H), 2.49-2.44 (m, 1H), 2.27-2.08 (m,2H), 1.45-1.33 (m, 2H), 1.24-1.14 (m, 2H), 0.80 (t, J=7.3 Hz, 3H).

Step C:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((pentanoyloxy)methyl)tetrahydrofuran-3-yltridecanoate. Tridecanoic acid (68.2 mg, 0.318 mmol) was dissolved inDMF (1 mL), DMAP (155 mg, 1.27 mmol) and EDC (244 mg, 1.27 mmol) wereadded. The resulting mixture was stirred for 2 h at RT. Then,((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylpentanoate (100 mg, 0.265 mmol) was added. The resulting mixture wasstirred for overnight at RT. LCMS indicated complete reaction. Thereaction mixture was quenched with water and extracted with EtOAc (3×).The organic phases were combined, washed with brine, dried over Na₂SO₄and concentrated under vacuum. The residue was subjected to preparativeTLC (MeOH:DCM=1:20) to give the desired product as a white solid (34 mg,22%). LCMS (ESI) m/z calcd for C₃₀H₄₄FN₅O₅: 573. Found: 574 (M+1)⁺. ¹HNMR (400 MHz, DMSO-d₆) δ 8.33 (s, 1H), 7.89 (br, 2H), 6.34 (t, J=6.7 Hz,1H), 5.70 (t, J=12.4 Hz, 1H), 4.40 (d, J=11.6 Hz, 1H), 4.21 (d, J=11.6Hz, 1H), 3.79 (s, 1H), 3.18-3.11 (m, 1H), 2.64-2.57 (m, 1H), 2.43-2.34(m, 2H), 2.32-2.18 (m, 2H), 1.59-1.54 (m, 2H), 1.47-1.39 (m, 2H),1.37-1.17 (m, 20H), 0.87-0.79 (m, 6H).

Example 20:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((pentanoyloxy)methyl)tetrahydrofuran-3-ylheptanoate

The title compound was prepared according to example 19, substitutingheptanoic acid for tridecanoic acid in step C. LCMS (ESI) m/z calcd forC₂₄H₃₂FN₅O₅: 489. Found: 490 (M+1)⁺. ¹H NMR (400 MHz, Methanol-d₄) δ8.18 (s, 1H), 6.38 (dd, J=7.2, 5.5 Hz, 1H), 5.82 (dd, J=7.3, 6.1 Hz,1H), 4.45 (d, J=11.8 Hz, 1H), 4.33 (d, J=11.7 Hz, 1H), 3.26 (s, 1H),3.22-3.15 (m, 1H), 2.75-2.69 (m, 1H), 2.43 (t, J=7.4 Hz, 2H), 2.32-2.24(m, 2H), 1.71-1.63 (m, 2H), 1.52-1.43 (m, 2H), 1.36-1.26 (m, 8H),0.92-0.86 (m, 6H).

Example 21:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((pentanoyloxy)methyl)tetrahydrofuran-3-ylundecanoate

The title compound was prepared according to example 19, substitutingundecanoic acid for tridecanoic acid in step C. LCMS (ESI) m/z calcd forC₂₈H₄₀FN₅O₅: 545. Found: 546 (M+1)⁺. ¹H NMR (400 MHz, Methanol-d₄) δ8.18 (s, 1H), 6.38 (dd, J=7.2, 5.5 Hz, 1H), 5.82 (dd, J=7.4, 6.1 Hz,1H), 4.45 (d, J=11.6 Hz, 1H), 4.33 (d, J=11.6 Hz, 1H), 3.25 (s, 1H),3.22-3.15 (m, 1H), 2.75-2.68 (m, 1H), 2.43 (t, J=7.4 Hz, 2H), 2.32-2.24(m, 2H), 1.71-1.64 (m, 2H), 1.53-1.49 (m, 2H), 1.39-1.24 (m, 16H),0.91-0.86 (m, 6H).

Example 22:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((pentanoyloxy)methyl)tetrahydrofuran-3-ylnonanoate

The title compound was prepared according to example 19, substitutingnonanoic acid for tridecanoic acid in step C. LCMS (ESI) m/z calcd forC₂₆H₃₆FN₅O₅: 517. Found: 518 (M+1)⁺. ¹H NMR (400 MHz, Methanol-d₄) δ8.18 (s, 1H), 6.38 (dd, J=7.2, 5.5 Hz, 1H), 5.82 (dd, J=7.4, 6.1 Hz,1H), 4.45 (d, J=11.8 Hz, 1H), 4.33 (d, J=11.8 Hz, 1H), 3.25 (s, 1H),3.22-3.15 (m, 1H), 2.75-2.68 (m, 1H), 2.43 (t, J=7.4 Hz, 2H), 2.33-2.22(m, 2H), 1.71-1.64 (m, 2H), 1.55-1.47 (m, 2H), 1.39-1.26 (m, 12H),0.92-0.85 (m, 6H).

Example 23:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((pentanoyloxy)methyl)tetrahydrofuran-3-ylpentanoate

The title compound was prepared according to example 19, substitutingpentanoic acid for tridecanoic acid in step C. LCMS (ESI) m/z calcd forC₂₂H₂₈FN₅O₅: 461. Found: 462 (M+1)⁺. ¹H NMR (400 MHz, Methanol-d₄) δ8.18 (s, 1H), 6.38 (dd, J=7.2, 5.6 Hz, 1H), 5.82 (dd, J=7.3, 6.0 Hz,1H), 4.45 (d, J=11.7 Hz, 1H), 4.33 (d, J=11.7 Hz, 1H), 3.26 (s, 1H),3.22-3.16 (m, 1H), 2.75-2.68 (m, 1H), 2.44 (t, J=7.4 Hz, 2H), 2.33-2.21(m, 2H), 1.69-1.62 (m, 2H), 1.56-1.47 (m, 2H), 1.45-1.36 (m, 2H),1.33-1.24 (m, 2H), 0.95 (t, J=7.3 Hz, 3H), 0.87 (t, J=7.3 Hz, 3H).

Example 24:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(propionyloxy)tetrahydrofuran-2-yl)methylpentanoate

The title compound was prepared according to example 19, substitutingpropanoic acid for tridecanoic acid in step C. LCMS (ESI) m/z calcd forC₂₀H₂₄FN₅O₅: 433. Found: 456 (M+23)⁺. ¹H NMR (400 MHz, Methanol-d₄) δ8.18 (s, 1H), 6.39 (dd, J=7.1, 5.8 Hz, 1H), 5.81 (dd, J=7.3, 5.8 Hz,1H), 4.45 (d, J=11.7 Hz, 1H), 4.33 (d, J=11.8 Hz, 1H), 3.26 (s, 1H),3.23-3.16 (m, 1H), 2.75-2.68 (m, 1H), 2.46 (dd, J=15.2, 7.6 Hz, 2H),2.36-2.20 (m, 2H), 1.56-1.45 (m, 2H), 1.34-1.24 (m, 2H), 1.18 (t, J=7.6Hz, 3H), 0.87 (t, J=7.3 Hz, 3H).

Example 25:((2R,3S,5R)-5-(6-butyramido-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylpentanoate

Step A:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methylpentanoate.((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylpentanoate (1.00 g, 2.65 mmol) was dissolved in DMF (20 mL), imidazole(0.541 g, 7.95 mmol) and tert-butylchlorodimethylsilane (1.20 g, 7.95mmol) was added at 0° C. The resulting mixture was stirred for 16 h atRT. LCMS indicated complete reaction. The reaction was diluted withwater and extracted with EA (3×). The organic phases were combined,washed with saturated brine, dried over Na₂SO₄ and concentrated undervacuum. The residue was subjected to flash chromatography (silica gel,40 g, EtOAc/PE=1:3) to give the desired product (1.0 g, 69%) as a whitesolid. LCMS (ESI) m/z calcd for C₂₃H₃₄FN₅O₄Si: 491. Found: 492 (M+1)⁺.¹H NMR (300 MHz, DMSO-d₆) δ 8.28 (s, 1H), 7.91 (br, 2H), 6.30-6.26 (m,1H), 4.99 (t, J=7.1 Hz, 1H), 4.35 (d, J=11.8 Hz, 1H), 4.10 (d, J=11.8Hz, 1H), 3.66 (s, 1H), 3.01-2.91 (m, 1H), 2.49-2.40 (m, 1H), 2.22-2.09(m, 2H), 1.40-1.38 (m, 2H), 1.24-1.16 (m, 2H), 0.92 (s, 9H), 0.80 (t,J=7.3 Hz, 3H), 0.15 (s, 6H).

Step B:((2R,3S,5R)-3-((tert-butyldimethylsilyl)oxy)-5-(6-butyramido-2-fluoro-9H-purin-9-yl)-2-ethynyltetrahydrofuran-2-yl)methylpentanoate.((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methylpentanoate (200 mg, 0.407 mmol) and DMAP (25.0 mg, 0.203 mmol) weredissolved in DCM (5 mL). TEA (0.227 mL, 1.63 mmol) was added at 0° C.Then a solution of butanoyl chloride (97.0 mg, 0.814 mmol) in DCM (2 mL)was added dropwise. Then the resulting mixture was stirred for 16 h atRT. LCMS indicated complete reaction. The reaction was quenched withwater, and extracted with EA (3×). The organic phases were combined,washed with saturated brine, dried over Na₂SO₄ and concentrated undervacuum. The residue was subjected to preparative TLC (100% EtOAc) togive the desired product (110 mg, 48%) as yellow oil. LCMS (ESI) m/zcalcd for C₂₇H₄₀FN₅O₅Si: 561. Found: 562 (M+1)⁺. ¹H NMR (400 MHz,Chloroform-d) δ 8.65 (s, 1H), 8.10 (s, 1H), 6.39-6.28 (m, 1H), 4.95-4.85(m, 1H), 4.46 (d, J=11.8 Hz, 1H), 4.28 (d, J=12.0 Hz, 1H), 3.51 (s, 1H),2.99-2.81 (m, 3H), 2.77-2.69 (m, 1H), 2.33-2.25 (m, 2H), 1.86-1.80 (m,2H), 1.61-1.56 (m, 2H), 1.37-1.30 (m, 2H), 1.07 (t, J=7.2 Hz, 3H), 0.96(s, 9H), 0.90 (t, J=7.3 Hz, 3H), 0.16 (d, J=4.4 Hz, 6H).

Step C:((2R,3S,5R)-5-(6-butyramido-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylpentanoate.((2R,3S,5R)-3-((tert-butyldimethylsilyl)oxy)-5-(6-butyramido-2-fluoro-9H-purin-9-yl)-2-ethynyltetrahydrofuran-2-yl)methylpentanoate (110 mg, 0.19 mmol) was dissolved in THF (10 mL), TBAF (0.38mL, 1M in THF, 0.38 mmol) was added, the resulting mixture was stirredfor 1 h at RT. LCMS indicated complete reaction. The reaction mixturewas concentrated under vacuum. Water was added, the mixture wasextracted with EtOAc (3×). The organic phases were combined, washed withbrine, dried over Na₂SO₄ and concentrated under vacuum. The residue wassubjected to preparative TLC (MeOH:DCM=1:10) followed by reverse phaseHPLC purification (C₁₈, MeCN/water with 0.1% formic acid)) to give thedesired product (25 mg, 41%). LCMS (ESI) m/z calcd for C₂₁H₂₆FN₅O₅: 447.Found: 448 (M+1)⁺. ¹H NMR (300 MHz, DMSO-d₆) δ 11.04 (s, 1H), 8.61 (s,1H), 6.35 (q, J=3.9 Hz, 1H), 5.85 (d, J=5.4 Hz, 1H), 4.75-4.71 (m, 1H),4.43 (d, J=11.7 Hz, 1H), 4.11 (d, J=11.7 Hz, 1H), 3.68 (s, 1H),2.91-2.83 (m, 1H), 2.58-2.51 (m, 3H), 2.28-2.07 (m, 2H), 1.62 (h, J=7.2Hz, 2H), 1.48-1.26 (m, 2H), 1.17 (h, J=7.2 Hz, 2H), 0.93 (t, J=7.2 Hz,3H), 0.78 (t, J=7.2 Hz, 3H).

Example 26:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-octanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methylpentanoate

The title compound was prepared according to example 25, substitutingoctanoyl chloride for butanoyl chloride in step B. LCMS (ESI) m/z calcdfor C₂₅H₃₄FN₅O₅: 503. Found: 504 (M+1)⁺. ¹HNMR (400 MHz, DMSO-d6) b11.04 (s, 1H), 8.60 (s, 1H), 6.36-6.33 (m, 1H), 5.85 (d, J=5.2 Hz, 1H),4.75-4.71 (m, 1H), 4.42 (d, J=11.6 Hz, 1H), 4.10 (d, J=11.6 Hz, 1H),3.68 (s, 1H), 2.89-2.83 (m, 1H), 2.55-2.49 (m, 3H), 2.21-2.18 (m, 1H),2.15-2.11 (m, 1H), 1.62-1.52 (m, 2H), 1.40-1.20 (m, 10H), 1.20-1.15 (m,2H), 0.90-0.81 (m, 3H), 0.77 (t, J=7.2 Hz, 3H).

Example 27:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methylpentanoate

The title compound was prepared according to example 25, substitutingtetradecanoyl chloride for butanoyl chloride in step B. LCMS (ESI) m/zcalcd for C₃₁H₄₆FN₅O₅: 587. Found: 588 (M+1)⁺. ¹H NMR (400 MHz,Methanol-d₄) δ 8.39 (s, 1H), 6.39 (dd, J=8.4, 3.6 Hz, 1H), 4.91-4.88 (m,1H), 4.46 (d, J=11.6 Hz, 1H), 4.24 (d, J=12.0 Hz, 1H), 3.19 (s, 1H),2.99-2.92 (m, 1H), 2.72-2.64 (m, 3H), 2.25-2.16 (m, 2H), 1.79-1.69 (m,2H), 1.52-1.19 (m, 24H), 0.89-0.82 (m, 6H).

Example 28:((2R,3S,5R)-5-(6-dodecanamido-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylpentanoate

The title compound was prepared according to example 25, substitutingdodecanoyl chloride for butanoyl chloride in step B. LCMS (ESI) m/zcalcd for C₂₉H₄₂FN₅O₅: 559. Found: 560 (M+1)⁺. ¹H NMR (400 MHz, DMSO-d₆)δ 11.04 (s, 1H), 8.60 (s, 1H), 6.35 (dd, J=8.0, 3.6 Hz, 1H), 5.85 (d,J=5.6 Hz, 1H), 4.75-4.71 (m, 1H), 4.43 (d, J=11.6 Hz, 1H), 4.11 (d,J=12.0 Hz, 1H), 3.68 (s, 1H), 2.86-2.82 (m, 1H), 2.57-2.49 (m, 3H),2.21-2.11 (m, 2H), 1.58-1.57 (m, 2H), 1.37-1.13 (m, 20H), 0.85 (t, J=6.8Hz, 3H), 0.78 (t, J=7.2 Hz, 3H).

Example 29:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(((2-propylpentanoyl)oxy)methyl)tetrahydrofuran-3-yldodecanoate

The title compound was prepared according to example 19, substituting2-propylpentanoyl chloride for pentanoyl chloride in step A anddodecanoic acid for tridecanoic acid in step C. LCMS (ESI) m/z calcd forC₃₂H₄₈FN₅O₅: 601. Found: 602 (M+1)⁺. ¹H NMR (400 MHz, Chloroform-d) δ8.05 (s, 1H), 6.42 (t, J=6.4 Hz, 1H), 5.99 (br, 2H), 5.63-5.60 (dd,J=7.1, 5.3 Hz, 1H), 4.47-4.39 (m, 2H), 3.00-2.95 (m, 1H), 2.77-2.72 (m,1H), 2.66 (s, 1H), 2.43-2.39 (m, 3H), 1.67-1.65 (m, 2H), 1.57-1.54 (m,2H), 1.42-1.41 (m, 2H), 1.26 (s, 20H), 0.88 (m, 9H).

Example 30:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-(butyryloxy)-2-ethynyltetrahydrofuran-2-yl)methyl2-propylpentanoate

The title compound was prepared according to example 19, substituting2-propylpentanoyl chloride for pentanoyl chloride in step A and butanoicacid for tridecanoic acid in step C. LCMS (ESI) m/z calcd forC₂₄H₃₂FN₅O₅: 489. Found: 490 (M+1)⁺. ¹H NMR (300 MHz, Methanol-d₄) δ8.18 (s, 1H), 6.42-6.38 (m, 1H), 5.86-5.81 (m, 1H), 4.37-4.32 (m, 2H),3.28-3.22 (m, 2H), 2.76-2.72 (m, 1H), 2.44-2.42 (m, 2H), 2.31-2.29 (m,1H), 1.75-1.67 (m, 2H), 1.51-1.45 (m, 2H), 1.39-1.14 (m, 6H), 1.00 (t,J=7.4 Hz, 3H), 0.83 (m, 6H).

Example 31:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-(butyryloxy)-2-ethynyltetrahydrofuran-2-yl)methyldecanoate

Step A:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-2-ethynyltetrahydrofuran-3-ylbutyrate. A suspension of(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-2-ethynyltetrahydrofuran-3-ol(800 mg, 1.505 mmol) in DCM (15 mL) was treated with butyric acid (0.275mL, 3.01 mmol), DMAP (184 mg, 1.51 mmol), EDC (865 mg, 4.51 mmol), DIEA(1.314 mL, 7.52 mmol), and stirred at RT for 1.5 h. The reaction wasconcentrated and purified by flash chromatography (silica gel, 0-100%EtOAc/DCM) to give the title compound (466 mg, 52%) as white solid. LCMS(ESI) m/z calcd for C₃₂H₃₆FN₅O₄Si: 601.3. Found: 602.3 (M+1)⁺. ¹H NMR(400 MHz, Chloroform-d) δ 7.96 (s, 1H), 7.71-7.63 (m, 4H), 7.48-7.33 (m,6H), 6.49 (dd, J=6.4, 7.2 Hz, 1H), 5.83 (dd, J=4.2, 6.8 Hz, 1H), 5.74(br s, 2H), 4.09-4.01 (m, 1H), 4.00-3.92 (m, 1H), 2.87 (td, J=7.1, 13.9Hz, 1H), 2.67 (ddd, J=4.1, 6.2, 13.8 Hz, 1H), 2.56 (s, 1H), 2.44-2.36(m, 2H), 1.79-1.68 (m, 2H), 1.10 (s, 9H), 1.00 (t, J=7.4 Hz, 3H).

Step B:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ylbutyrate. An ice cold solution of(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-2-ethynyltetrahydrofuran-3-ylbutyrate (665 mg, 1.105 mmol) in THF (22 mL) was treated with TBAF (1Min THF) (1.66 mL, 1.66 mmol) and stirred at 0° C. for 2 h. The reactionwas quenched with AcOH (˜1 mL), diluted with water, and extracted withEtOAc. The combined organics were washed with brine 4×, dried overNa₂SO₄, filtered, and concentrated. Purification by flash chromatography[silica gel, 0-100% (3:1 EtOAc:EtOH)/hexanes] afforded the titlecompound (297 mg, 70%) as white solid. LCMS (ESI) m/z calcd forC₁₆H₁₈FN₅O₄: 363.1. Found: 364.1 (M+1)⁺. ¹H NMR (400 MHz, DMSO-d₆) δ8.33 (s, 1H), 7.87 (br s, 2H), 6.32 (dd, J=6.2, 7.9 Hz, 1H), 5.60 (dd,J=3.2, 6.6 Hz, 1H), 5.54 (dd, J=5.5, 6.9 Hz, 1H), 3.75-3.56 (m, 3H),3.00 (ddd, J=6.8, 7.7, 14.1 Hz, 1H), 2.57-2.52 (m, 1H), 2.43-2.33 (m,2H), 1.62 (sxt, J=7.3 Hz, 2H), 0.94 (t, J=7.4 Hz, 3H).

Step C((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-(butyryloxy)-2-ethynyltetrahydrofuran-2-yl)methyldecanoate. A suspension of(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ylbutyrate (248 mg, 0.683 mmol) in DCM (6.8 mL) was treated with decanoicacid (141 mg, 0.819 mmol), DMAP (83 mg, 0.683 mmol), EDC (236 mg, 1.229mmol), DIEA (0.596 mL, 3.41 mmol), and stirred at RT for 3.5 h. Thereaction was diluted with water and extracted with DCM. The DCM solutionwas washed with sat. NaHCO₃, brine, dried over Na₂SO₄, filtered, andconcentrated. Purification by flash chromatography (silica gel, 0-100%EtOAc/DCM) afforded the title compound (261 mg, 70%) as white solid.LCMS (ESI) m/z calcd for C₂₆H₃₆FN₅O₅: 517.3. Found: 518.8 (M+1)⁺. ¹H NMR(400 MHz, DMSO-d₆) δ 8.31 (s, 1H), 7.84 (br s, 2H), 6.34 (t, J=6.8 Hz,1H), 5.69 (dd, J=5.2, 6.9 Hz, 1H), 4.40 (d, J=11.7 Hz, 1H), 4.23 (d,J=11.7 Hz, 1H), 3.76 (s, 1H), 3.15 (td, J=6.9, 13.8 Hz, 1H), 2.61 (ddd,J=5.2, 6.9, 14.1 Hz, 1H), 2.38 (dt, J=2.9, 7.3 Hz, 2H), 2.34-2.15 (m,2H), 1.62 (sxt, J=7.3 Hz, 2H), 1.53-1.38 (m, 2H), 1.31-1.12 (m, 12H),0.93 (t, J=7.4 Hz, 3H), 0.89-0.80 (m, 3H).

Example 32:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(heptanoyloxy)tetrahydrofuran-2-yl)methylbenzoate

The title compound was prepared according to example 31, substitutingbenzoic acid for butyric acid in step A and heptanoic acid for decanoicacid in step C. LCMS (ESI) m/z calcd for C₂₆H₂₈FN₅O₅: 509.2. Found:510.5 (M+1)⁺. ¹H NMR (400 MHz, Chloroform-d) b 8.06-7.99 (m, 2H), 7.85(s, 1H), 7.61-7.54 (m, 1H), 7.49-7.41 (m, 2H), 6.42 (t, J=6.6 Hz, 1H),5.84 (dd, J=5.5, 7.2 Hz, 1H), 5.77 (br s, 2H), 4.80 (d, J=11.9 Hz, 1H),4.62 (d, J=11.9 Hz, 1H), 3.11 (ddd, J=6.4, 7.4, 13.8 Hz, 1H), 2.81-2.67(m, 2H), 2.44 (t, J=7.5 Hz, 2H), 1.76-1.64 (m, 2H), 1.44-1.28 (m, 6H),0.96-0.87 (m, 3H).

Example 33:(2R,3aS,20aR)-2-(6-amino-2-fluoro-9H-purin-9-yl)-20a-ethynylhexadecahydro-2H-furo[3,2-b][1,5]dioxacyclononadecine-5,18-dione

Step A:1-((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl)14-tert-butyl tetradecanedioate. To a stirred solution of(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-2-ethynyltetrahydrofuran-3-ol(146 mg, 0.275 mmol), 14-(tert-butoxy)-14-oxotetradecanoic acid (130 mg,0.412 mmol) and DMAP (33.5 mg, 0.275 mmol) in DCM (1.5 mL) at ambienttemperature was added EDC (158 mg, 0.824 mmol), followed by DIPEA (0.240mL, 1.373 mmol) and the mixture was allowed to stir overnight. Themixture was concentrated and then purified on silica gel (0-50%DCM/EtOAc) to afford the title compound (187 mg, 82%) as a colorlessresidue. LCMS (ESI) m/z calcd for C₄₆H₆₂FN₅O₆Si: 827.5. Found: 828.7(M+1)⁺. ¹H NMR (400 MHz, Chloroform-d) δ 7.96 (s, 1H), 7.70-7.63 (m,4H), 7.47-7.35 (m, 6H), 6.49 (dd, J=6.2, 7.2 Hz, 1H), 5.90-5.74 (m, 3H),4.05 (d, J=11.2 Hz, 1H), 3.96 (d, J=11.2 Hz, 1H), 2.90-2.81 (m, 1H),2.70-2.63 (m, 1H), 2.56 (s, 1H), 2.41 (t, J=7.5 Hz, 2H), 2.21 (t, J=7.5Hz, 2H), 1.75-1.52 (m, 4H), 1.45 (s, 9H), 1.40-1.22 (m, 16H), 1.10 (s,9H).

Step B:1-((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-yl)14-tert-butyl tetradecanedioate. To a solution of1-((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl)14-(tert-butyl) tetradecanedioate (185 mg, 0.223 mmol) in THF (2.0 mL)was added TBAF, 1M solution in THF (0.335 mL, 0.335 mmol) and themixture was allowed to stir at ambient temperature for 60 minutes. AcOH(335 uL) was added and the mixture was concentrated. The residue waspurified on silica gel (0-10% DCM/MeOH) to afford the title compound(101 mg, 77%) as a white solid. LCMS (ESI) m/z calcd for C₃₀H₄₄FN₅O₆:589.3. Found: 590.6 (M+1)⁺. ¹H NMR (400 MHz, Chloroform-d) b 7.84 (s,1H), 6.34 (dd, J=5.5, 9.3 Hz, 1H), 5.97-5.72 (m, 3H), 5.41 (dd, J=3.3,11.4 Hz, 1H), 4.07-4.01 (m, 1H), 3.98-3.90 (m, 1H), 3.21 (ddd, J=6.3,9.3, 13.7 Hz, 1H), 2.62 (s, 1H), 2.50-2.38 (m, 3H), 2.21 (t, J=7.5 Hz,2H), 1.74-1.65 (m, 2H), 1.63-1.52 (m, 2H), 1.45 (s, 9H), 1.41-1.23 (m,16H).

Step C:14-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-yl)oxy)-14-oxotetradecanoicacid. To a solution of1-((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-yl)14-(tert-butyl) tetradecanedioate (101 mg, 0.171 mmol) in DCM (3.0 mL)at ambient temperature was added TFA (50 μl, 0.649 mmol) and the mixturewas stirred for 25 minutes. LCMS indicated mostly starting materialremained. Additional TFA (50 uL) was added and the mixture was stirredfor six hours. The mixture was concentrated and then purified on silicagel (0-10% DCM/MeOH) to afford the title compound (56 mg, 61%) as awhite solid. LCMS (ESI) m/z calcd for C₂₆H₃₆FN₅O₆: 533.3. Found: 534.5(M+1)⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 12.03-11.75 (m, 1H), 8.33 (s, 1H),7.97-7.74 (m, 2H), 6.31 (dd, J=6.4, 7.9 Hz, 1H), 5.67-5.41 (m, 2H),3.79-3.55 (m, 3H), 3.04-2.94 (m, 1H), 2.57-2.46 (m, 1H, overlapping DMSOpeak), 2.41-2.34 (m, 2H), 2.17 (t, J=7.4 Hz, 2H), 1.65-1.40 (m, 4H),1.37-1.10 (m, 16H).

Step D:(2R,3aS,20aR)-2-(6-amino-2-fluoro-9H-purin-9-yl)-20a-ethynylhexadecahydro-2H-furo[3,2-b][1,5]dioxacyclononadecine-5,18-dione.To a solution of14-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-yl)oxy)-14-oxotetradecanoicacid (56 mg, 0.105 mmol) in DCM (6.5 mL) at ambient temperature wassequentially added DMAP (12.82 mg, 0.105 mmol), DIPEA (0.092 mL, 0.525mmol) and EDC (60.4 mg, 0.315 mmol) and the mixture was allowed to stirfor 3 days. The mixture was concentrated in the presence of Celite.Silica gel chromatography (0-50% DCM/EtOAc) afforded the title compound(4.6 mg, 8.5%) as a colorless residue. LCMS (ESI) m/z calcd forC₂₆H₃₄FN₅O₅: 515.3. Found: 516.8 (M+1)⁺. ¹H NMR (400 MHz, Chloroform-d)δ 7.92 (s, 1H), 6.39 (dd, J=5.5, 6.9 Hz, 1H), 5.87 (br s, 2H), 5.76-5.71(m, 1H), 4.48 (d, J=11.9 Hz, 1H), 4.40 (d, J=11.7 Hz, 1H), 3.15-3.06 (m,1H), 2.79-2.70 (m, 1H), 2.66 (s, 1H), 2.48-2.35 (m, 4H), 1.79-1.61 (m,4H), 1.48-1.18 (m, 16H).

Example 34:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylheptoanoylglycinate

Step A:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methyl2-((tert-butoxycarbonyl)amino)acetate. To a stirred solution of((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methanol(31 mg, 0.037 mmol), (tert-butoxycarbonyl)glycine (12.96 mg, 0.074 mmol)and DMAP (4.52 mg, 0.037 mmol) in DCM (0.6 mL) at ambient temperaturewas added EDC (21.28 mg, 0.111 mmol), followed by DIPEA (0.032 mL, 0.185mmol). The mixture was allowed to stir for 24 hours. Additional(tert-butoxycarbonyl)glycine (12.96 mg, 0.074 mmol), DMAP (4.52 mg,0.037 mmol), EDC (21.28 mg, 0.111 mmol) and DIPEA (0.032 mL, 0.185 mmol)were added and the mixture was stirred overnight. The mixture wasconcentrated and then purified on silica gel (0-50% DCM/EtOAc) to affordthe title compound (16 mg, 44%) as a colorless residue. LCMS (ESI) m/zcalcd for C₅₉H₅₅FN₆O₈: 995.1. Found: 996.3 (M+1)⁺.

Step B:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl2-aminoacetate. To a solution of((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methyl2-((tert-butoxycarbonyl)amino)acetate (16 mg, 0.016 mmol) in DCM (0.8mL) was added TFA (0.2 ml). The resulting orange solution was stirredovernight, then concentrated and used in the next step without furtherpurification. LCMS (ESI) m/z calcd for C₁₄H₁₅FN₆O₄: 350.1. Found: 351.7(M+1)⁺.

Step C:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylheptanoylulycinate. To a solution of the crude amine product from theprevious step in DMF (0.6 mL) was added heptanoic acid (6.81 μl, 0.048mmol), DIPEA (0.014 mL, 0.080 mmol) and finally HATU (12.17 mg, 0.032mmol). The mixture was stirred for 20 minutes then purified directly byRP-HPLC (C18, 10-100% MeCN/water with 0.1% FA) to afford the titlecompound (4.6 mg, 27% over two steps) as a white residue. LCMS (ESI) m/zcalcd for C₂₁H₂₇FN₆O₅: 462.2. Found: 463.8 (M+1)⁺. ¹H NMR (400 MHz,Methanol-d₄) δ 8.18 (s, 1H), 6.32 (dd, J=3.9, 7.7 Hz, 1H), 4.85-4.77 (m,1H, overlapping water signal), 4.50 (d, J=11.7 Hz, 1H), 4.37 (d, J=11.9Hz, 1H), 3.96 (d, J=17.6 Hz, 1H), 3.84 d, J=17.6 Hz, 1H), 3.16 (s, 1H),2.90-2.83 (m, 1H), 2.69-2.61 (m, 1H), 2.26-2.16 (m, 2H), 1.66-1.51 (m,2H), 1.39-1.20 (m, 6H), 0.94-0.84 (m, 3H).

Example 35:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyltetradecanoylglycinate

Step A: tert-butyl 2-tetradecanamidoacetate. To a solution oftetradecanoic acid (200 mg, 0.876 mmol), tert-butyl glycinate,hydrochloride (176 mg, 1.051 mmol) and DIPEA (0.459 mL, 2.63 mmol) inDMF (5 mL) was added HATU (533 mg, 1.401 mmol) and the mixture wasallowed to stir at ambient temperature for 3 hours. Water was added andthe mixture was extracted with EtOAc. The extracts were washed with 1NHCl, then sat'd NaHCO₃ at which point an emulsion formed. Brine wasadded and the mixture was filtered over Celite and the layers wereseparated. The organic phase was further washed with water, then brine,dried over Na₂SO₄, filtered and concentrated to afford the titlecompound (98 mg, 33%) as a white solid. ¹H NMR (400 MHz, DMSO-d₆)δ=8.10-8.03 (m, 1H), 3.66 (d, J=6.0 Hz, 2H), 2.08 (t, J=7.4 Hz, 2H),1.53-1.43 (m, 2H), 1.39 (s, 9H), 1.23 (s, 20H), 0.89-0.81 (m, 3H).

Step B: 2-tetradecanamidoacetic acid. A solution of tert-butyltetradecanoylglycinate (98 mg, 0.287 mmol) in DCM (0.8 mL) was treatedwith TFA (0.2 mL, 2.60 mmol) and the mixture was stirred at ambienttemperature for 2 hours. Additional TFA (0.2 mL, 2.60 mmol) was addedand stirring at ambient temperature was continued overnight. The mixturewas concentrated to give a sticky brown solid which was used withoutfurther purification. ¹H NMR (400 MHz, DMSO-d₆) δ 12.40 (br. s, 1H),8.08-8.02 (m, 1H), 3.72 (d, J=6.0 Hz, 2H), 2.10 (t, J=7.4 Hz, 2H),1.56-1.42 (m, 2H), 1.34-1.15 (m, 20H), 0.91-0.80 (m, 3H).

Steps C and D:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyltetradecanoylqlycinate. The title compound was prepared from((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methanoland 2-tetradecanamidoacetic acid according to example 1, steps E and F.LCMS (ESI) m/z calcd for C₂₈H₄₁FN₆O₅: 560.3. Found: 561.5 (M+1)⁺. ¹H NMR(400 MHz, Methanol-d₄) δ 8.18 (s, 1H), 6.33 (dd, J=3.9, 7.7 Hz, 1H),4.87-4.74 (m, 1H, overlapping H₂O peak), 4.50 (d, J=11.9 Hz, 1H), 4.38(d, J=11.7 Hz, 1H), 3.97 (d, J=17.6 Hz, 1H), 3.84 (d, J=17.6 Hz, 1H),3.16 (s, 1H), 2.91-2.83 (m, 1H), 2.70-2.61 (m, 1H), 2.27-2.15 (m, 2H),1.65-1.52 (m, 2H), 1.28 (s, 20H), 0.93-0.85 (m, 3H).

Example 36:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylheptanoyl-L-alaninate

Step A:(S)-((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)-amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methyl2-((tert-butoxvcarbonyl)amino)propanoate. To a stirred solution of((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methanol(39 mg, 0.047 mmol), (tert-butoxycarbonyl)-L-alanine (17.61 mg, 0.093mmol) and DMAP (5.69 mg, 0.047 mmol) in DCM (0.5 mL) at ambienttemperature was added EDC (26.8 mg, 0.140 mmol), followed by DIPEA(0.041 mL, 0.233 mmol). The mixture was allowed to stir at ambienttemperature overnight. The mixture was concentrated and then purified onsilica gel (0-50% hexanes/EtOAc) to afford the title compound (41 mg,87%) as a colorless residue (41 mg). LCMS (ESI) m/z calcd forC₆₀H₅₇FN₆O₈: 1008.4. Found: 1009.6 (M+1)⁺.

Step B:(S)-((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetra-hydrofuran-2-yl)methyl2-aminopropanoate. To a solution of(S)-((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methyl2-((tert-butoxycarbonyl)-amino)propanoate (41 mg, 0.041 mmol) in DCM(0.8 mL) was added TFA (0.2 ml). The resulting orange solution wasstirred for two hours, then concentrated and used in the next stepwithout further purification. LCMS (ESI) m/z calcd for C₁₅H₁₇FN₆O₄:364.1. Found: 365.1 (M+1)⁺.

Step C:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetra-hydrofuran-2-yl)methylheptanoyl-L-alaninate. To a mixture of the crude product from theprevious step in DMF (0.500 mL) was added heptanoic acid (6.06 mg, 0.047mmol), DIPEA (0.024 mL, 0.140 mmol) and finally HATU (28.3 mg, 0.074mmol) at ambient temperature and the mixture was allowed to stir for 30minutes. Sat'd NaHCO₃ was added and the mixture was extracted withEtOAc. The combined extracts were washed with water, then brine, driedover Na₂SO₄, filtered and concentrated to a pale brown. The residue wasdissolved in DCM (0.8 mL), treated with formic acid (0.2 mL), followedby triethylsilane (0.015 mL, 0.093 mmol) and the mixture was stirred atambient temperature for 30 minutes. The mixture was concentrated andthen purified on silica gel (0-10% DCM/MeOH) to afford the titlecompound (10.2 mg, 51% over two steps) as a colorless residue. LCMS(ESI) m/z calcd for C₂₂H₂₉FN₆O₆: 476.2. Found: 477.2 (M+1)⁺. ¹H NMR (400MHz, DMSO-d₆) δ 8.26 (s, 1H), 8.14-8.09 (m, 1H), 7.80 (br s, 2H), 6.25(dd, J=5.0, 7.6 Hz, 1H), 5.77-5.74 (m, 1H), 4.68-4.52 (m, 1H), 4.32 (d,J=11.4 Hz, 1H), 4.24-4.15 (m, 2H), 3.59 (s, 1H), 2.84-2.75 (m, 1H),2.53-2.40 (m, 1H, overlapping DMSO peak), 2.09-2.02 (m, 2H), 1.52-1.35(m, 2H), 1.31-1.09 (m, 9H), 0.89-0.74 (m, 3H).

Example 37:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-(butyryloxy)-2-ethynyltetrahydrofuran-2-yl)methyltetradecanoate

The title compound was prepared according to example 31, substitutingtetradecanoic acid for decanoic acid in step C. LCMS (ESI) m/z calcd forC₃₀H₄₄FN₅O₅: 573. Found: 574 (M+1)⁺. ¹H NMR (400 MHz, Chloroform-d) δ7.95 (s, 1H), 6.42 (t, J=6.4 Hz, 1H), 5.80 (br s, 2H), 5.68 (dd, J=6.9,5.5 Hz, 1H), 4.51 (d, J=11.9 Hz, 1H), 4.41 (d, J=11.9 Hz, 1H), 3.03(ddd, J=13.8, 7.5, 6.2 Hz, 1H), 2.67-2.76 (m, 2H), 2.32-2.47 (m, 4H),1.57-1.81 (m, 4H), 1.24-1.34 (m, 20H), 1.02 (t, J=7.39 Hz, 3H),0.87-0.94 (m, 3H).

Example 38:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methyltetradecanoate

Step A: hexyl (4-nitrophenyl) carbonate. A mixture of 4-nitrophenylcarbonochloridate (2.00 g, 9.92 mmol) and hexan-1-ol (1.24 ml, 9.92mmol) in DCM (20 mL) was treated with TEA (2.08 ml, 14.9 mmol) and themixture was stirred at RT for 1 h. The reaction was concentrated andused in the next step without further purification.

Step B:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-2-ethynyltetrahydrofuran-3-ylhexyl carbonate. A mixture of(2R,3S,5R)-5-(6-amino-2-fluoro-4,5-dihydro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-2-ethynyltetrahydrofuran-3-ol(300 mg, 0.562 mmol) and DMAP (68.7 mg, 0.562 mmol) in DCM (6.0 mL) wastreated with TEA (0.235 mL, 1.69 mmol) followed by hexyl (4-nitrophenyl)carbonate (300 mg, 1.12 mmol) and the mixture was stirred at RT for 2days. Concentrated and purified by flash chromatography (silica gel,0-10% MeOH/DCM) to give the desired product as a white foam. (332 mg,90%). LCMS (ESI) m/z calcd for C₃₅H₄₂FN₅O₅Si: 659. Found: 660 (M+1)⁺. ¹HNMR (400 MHz, Chloroform-d) δ 8.00 (s, 1H), 7.64-7.73 (m, 4H), 7.35-7.47(m, 6H), 6.62 (s, 2H), 6.51 (t, J=6.7 Hz, 1H), 5.75 (dd, J=6.8, 4.4 Hz,1H), 4.18-4.29 (m, 2H), 4.12 (d, J=11.2 Hz, 1H), 4.02 (d, J=11.0 Hz,1H), 2.98 (dt, J=13.9, 7.0 Hz, 1H), 2.81 (ddd, J=13.8, 6.4, 4.4 Hz, 1H),2.64 (s, 1H), 1.68-1.79 (m, 2H), 1.26-1.46 (m, 6H), 1.06-1.16 (m, 9H),0.87-0.99 (m, 3H).

Step C:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetra-hydrofuran-3-ylhexyl carbonate. To a solution of(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl hexyl carbonate (332 mg, 0.503mmol) in THF (8.0 mL) was added TBAF (1M, THF) (0.843 mL, 0.843 mmol)and the mixture was stirred at RT for 1 h. The mixture was concentratedand the residue purified by flash chromatography (silica gel, 0-10%MeOH/DCM) to give the desired product as a white solid. (203 mg, 96%).LCMS (ESI) m/z calcd for C₁₉H₂₄FN₅O₅: 421. Found: 422 (M+1)⁺. ¹H NMR(400 MHz, Chloroform-d) δ 7.82 (s, 1H), 6.37 (dd, J=9.4, 5.4 Hz, 1H),6.02 (br s, 2H), 5.66 (dd, J=6.2, 1.2 Hz, 1H), 4.18-4.29 (m, 2H), 4.07(d, J=12.4 Hz, 1H), 3.97 (br d, J=12.6 Hz, 1H), 3.25 (ddd, J=13.9, 9.5,6.2 Hz, 1H), 2.66 (s, 1H), 2.57 (ddd, J=14.0, 5.6, 1.4 Hz, 1H),1.66-1.85 (m, 3H), 1.27-1.47 (m, 6H), 0.88-0.97 (m, 3H).

Step D:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)-ox)tetrahydrofuran-2-yl)methyltetradecanoate. To a stirred solution of(2R,3S)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ylhexyl carbonate (50.0 mg, 0.119 mmol), EDC (45.5 mg, 0.237 mmol) andDMAP (14.5 mg, 0.119 mmol) in DCM (3.0 mL) was added tetradecanoic acid(32.5 mg, 0.142 mmol) followed by DIEA (0.083 mL, 0.48 mmol) and themixture was stirred at RT for 16 h. The reaction mixture wasconcentrated and purified by flash chromatography (silica gel, 0-10%MeOH/DCM) to afford the product as a white solid. (58 mg, 77% yield).LCMS (ESI) m/z calcd for C₃₃H₅₀FN₅O₆: 631. Found: 632 (M+1)⁺. ¹H NMR(400 MHz, Chloroform-d) δ 7.91 (s, 1H), 6.50 (s, 2H), 6.39 (t, J=6.4 Hz,1H), 5.60 (dd, J=7.0, 5.6 Hz, 1H), 4.55 (d, J=11.9 Hz, 1H), 4.41 (d,J=11.9 Hz, 1H), 4.21 (m, 2H), 3.14 (dt, J=13.4, 7.0 Hz, 1H), 2.80 (ddd,J=13.9, 6.7, 5.6 Hz, 1H), 2.71 (s, 1H), 2.28-2.42 (m, 2H), 1.55-1.76 (m,4H), 1.28-1.44 (m, 10H), 1.25 (br s, 16H), 0.83-0.96 (m, 6H).

Example 39:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(((2-propylpentanoyl)oxy)methyl)tetrahydrofuran-3-ylheptanoate

The title compound was prepared according to example 19, substituting2-propylpentanoyl chloride for pentanoyl chloride in step A andheptanoic acid for tridecanoic acid in step C. LCMS (ESI) m/z calcd forC₂₇H₃₈FN₅O₅: 531. Found: 532 (M+1)⁺. ¹H NMR (400 MHz, Chloroform-d) δ7.96 (s, 1H), 6.43 (t, J=6.4 Hz, 1H), 5.84 (br s, 2H), 5.65 (dd, J=7.0,5.4 Hz, 1H), 4.41-4.49 (m, 2H), 3.01 (ddd, J=13.7, 7.2, 6.3 Hz, 1H),2.66-2.77 (m, 2H), 2.39-2.48 (m, 3H), 1.55-1.78 (m, 4H), 1.26-1.50 (m,12H), 0.86-0.94 (m, 9H).

Example 40:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyltetradecanoyl-L-alaninate

Step 1: tetradecanoyl-L-alanine. To a solution of L-alanine (1.6 g,17.96 mmol), sodium bicarbonate (3.02 g, 35.9 mmol) in THF (40.0mL)/Water (40 mL) at 20° C. was added tetradecanoyl chloride (2.216 g,8.98 mmol) dropwise over 15 min. The reaction mixture was stirred atr.t. for 5 h. LCMS indicated completion of reaction. The reactionmixture was diluted with water (30 ml), and extracted with EtOAc (50ml*2). The aqueous phase was acidified with 1 M hydrochloric acid. Theresulting solid was filtered through a Buchner funnel, rinsed with water(40 mL), and collected to give the crude tetradecanoyl-L-alanine as awhite solid (800 mg, 27%). LCMS (ESI) m/z calcd for C₁₇H₃₃NO₃:299.Found: 300 (M+H)⁺. ¹HNMR (400 MHz, DMSO-d₆) δ8.02-7.91 (m, 1H),4.16-4.05 (m, 1H), 2.05 (t, J=7.2 Hz, 2H), 1.48-1.41 (m, 2H), 1.26-1.17(m, 23H), 0.83 (t, J=6.8 Hz, 3H).

Step 2:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methyl-tetradecanoyl-L-alaninate.Tetradecanoyl-L-alanine (1 g, 3.34 mmol) was dissolved in DCM (30 mL)and added EDC (3.20 g, 16.70 mmol) and DMAP (2.040 g, 16.70 mmol). Theresulting mixture was stirred for 40 min at room temperature. Then,((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphen-yl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethyl-oxy)tetrahydro-furan-2-yl)methanol(1.399 g, 1.670 mmol) was added and the resulting mixture was stirredovernight at room temperature. LCMS indicated completion of reaction.The reaction mixture was concentrated to dryness under vacuum to affordthe crude product. The crude product was pre-absorbed on silica andpurified on a silica column (120 g) using a 0%-60% pet. ether-EtOAcsolvent gradient over 60 mins, Flow rate: 70 mL/min. The appropriatefractions were identified by UV absorbance (254 nm), combined andevaporated under vacuum to give the title product as a white solid (800mg, 19%). LCMS (ESI) m/z calcd for C₆₉H₇₅FN₆O₇: 1119, Found: 1120(M+H)⁺. ¹HNMR (400 MHz, CDCl₃) δ7.55-7.49 (m, 4H), 7.42-7.37 (m, 2H),7.33-7.18 (m, 18H), 6.80 (t, J=12 Hz, 4H) 5.90 (dd, J=24.4, 7.6 Hz, 1H),4.47-4.28 (m, 2H), 4.15-4.06 (m, 4H), 3.78 (s, 3H), 3.74 (d, J=8.0 Hz,1H), 2.82 (d, J=8.0 Hz, 1H), 2.05 (s, 4H), 1.29-1.24 (m, 22H), 1.19-1.13(m, 2H), 1.11-1.05 (m, 2H), 0.85 (t, J=6.4 Hz, 3H).

Step 3:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydro-furan-2-yl)methyltetradecanoyl-L-alaninate. To a stirred solution of((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)di-phenylmethoxy)tetrahydrofuran-2-yl)methyltetradecanoyl-L-alaninate (800 mg, 0.715 mmol) in DCM (3 mL) at 15° C.was added TFA (300 μL, 3.89 mmol) dropwise. The reaction mixture wasstirred at 15° C. for 1 h. LCMS indicated completion of reaction. Thereaction mixture was diluted with NaHCO₃ (30 ml), and extracted with DCM(15 ml*2). The combined organic phases were washed with brine (15 ml).The organic layers were dried over anhydrous sodium sulphate andconcentrated under reduced pressure to afford crude product as a whitesolid. The sample was pre-absorbed on silica and purified on a silicacolumn (80 g) using a 0%-10% DCM-MeOH solvent gradient over 30 mins,Flow rate: 45 mL/min. The appropriate fractions were identified by UVabsorbance (254 nm), combined and evaporated in vacuo to give((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydro-furan-2-yl)methyltetradecanoyl-L-alaninate (189 mg, 46%) as a white solid. LCMS (ESI) m/zcalcd for C₂₉H₄₃FN₆O₅:574. Found: 575 (M+H)⁺. ¹HNMR (400 MHz, CDCl₃) δ7.95 (s, 1H), 6.32 (dd, J=7.2, 4.8 Hz, 1H), 6.05-6.07 (m, 3H), 4.75 (t,J=6.4 Hz, 1H), 4.61 (d, J=11.6 Hz, 1H), 4.52 (t, J=6.8 Hz, 1H), 4.42 (d,J=11.6, 1H), 3.31 (s, 1H), 2.98-3.02 (m, 1H), 2.79 (s, 1H), 2.63-2.70(m, 1H), 2.21 (t, J=7.6 Hz, 2H), 1.58-1.63 (m, 2H), 1.40 (d, J=7.2 Hz,3H) 1.24-1.31 (m, 20H), 0.83-0.89 (m, 3H).

Example 41:(2R,3S,5R)-2-(acetoxymethyl)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyltetrahydrofuran-3-ylheptanoate

The title compound was isolated as a by product from example 32preparation. This product was formed due to the presence of acetic acidin the starting material. ¹HNMR (400 MHz, CHLOROFORM-d) δ ppm 7.92 (s,1H), 6.39 (t, J=6.4 Hz, 1H), 5.77 (br s, 2H), 5.69 (dd, J=5.5, 7.2 Hz,1H), 4.52 (d, J=12.2 Hz, 1H), 4.38 (d, J=12.2 Hz, 1H), 3.04 (ddd, J=6.1,7.4, 13.7 Hz, 1H), 2.76-2.63 (m, 2H), 2.42 (t, J=7.5 Hz, 2H), 2.12 (s,3H), 1.75-1.63 (m, 2H), 1.41-1.28 (m, 6H), 0.95-0.87 (m, 3H). LCMS (m/z)ES⁺=448.2 (M+1), ES⁻=446.4 (M−1).

Example 42:((2R,3S,5R)-5-(6-decanamido-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylheptanoate

Step 1:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methylheptanoate. To a suspension of((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methanol (390 mg, 0.861 mmol) in DCM(25 mL) was added heptanoic acid (0.271 mL, 1.913 mmol) followed by DMAP(117 mg, 0.958 mmol), EDC (550 mg, 2.87 mmol) and DIEA (0.835 mL, 4.78mmol) and the mixture was stirred at ambient temperature for 1.5 h. Themixture was diluted with dichloromethane and washed with water. Theorganic phases were dried (Na₂SO₄), concentrated and purified on silicagel (Biotage Isolera 1, 24 g, Gold column, EtOAc/hexanes 0-25% then50-100%, bis-silyl impurity eluted at 25%, product eluted at 50-100%) toprovide((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methylheptanoate (360 mg, 0.686 mmol, 80% yield) as an off-white solid. ¹HNMR(400 MHz, CHLOROFORM-d) δ ppm 7.87 (s, 1H), 6.27 (dd, J=7.75, 3.70 Hz,1H), 5.82 (br s, 2H), 4.96 (t, J=7.39 Hz, 1H), 4.48 (d, J=11.92 Hz, 1H),4.26 (d, J=11.92 Hz, 1H), 2.84-2.98 (m, 1H), 2.58-2.72 (m, 2H), 2.28(td, J=7.57, 3.46 Hz, 2H), 1.50-1.61 (m, 2H), 1.19-1.42 (m, 6H),0.93-1.03 (m, 9H), 0.81-0.92 (m, 3H), 0.17 (d, J=4.29 Hz, 6H). LCMS(M+1)=520.7.

Step 2:((2R,3S,5R)-3-((tert-butyldimethylsilyl)oxy)-5-(6-decanamido-2-fluoro-9H-purin-9-vi)-2-ethynyltetrahydrofuran-2-yl)methylheptanoate. To a solution of((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methylheptanoate (280 mg, 0.539 mmol) in DCM (7 mL) was added DIEA (0.565 mL,3.23 mmol) and DMAP (65.8 mg, 0.539 mmol). The mixture was cooled to 0°C. and decanoyl chloride (0.335 mL, 1.616 mmol) was added dropwise.Stirring at 0° C. continued for 30 min [LCMS showed only s.m.] and thenat ambient temperature for 1.5 h. The mixture was diluted withdichloromethane and washed with saturated NaHCO₃/water and the organicphases were dried (Na₂SO₄), concentrated, and purified on silica gel(Biotage Isolera 1, 24 g column, EtOAc/hexanes 0-10-30-100%, 50-1-1eluted at 30%, 50-2 at 30-35%, recovered s.m. eluted at 70%) to provide((2R,3S,5R)-3-((tert-butyldimethylsilyl)oxy)-5-(6-decanamido-2-fluoro-9H-purin-9-yl)-2-ethynyltetrahydrofuran-2-yl)methylheptanoate (143.5 mg, 0.213 mmol, 39.5% yield) as a yellowish oil andrecovered starting material. LCMS (M+1)=675.

Step 3:((2R,3S,5R)-5-(6-decanamido-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl)methylheptanoate. To a solution of((2R,3S,5R)-3-((tert-butyl-dimethylsilyl)oxy)-5-(6-decanamido-2-fluoro-9H-purin-9-yl)-2-ethynyltetrahydrofuran-2-yl)methylheptanoate (138 mg, 0.205 mmol) in THF (2.5 mL) was added a solution ofTBAF (0.410 mL, 0.410 mmol) (1 M/THF) containing acetic acid (0.023 mL,0.410 mmol) and the mixture was stirred at ambient temperature for 5 h.The mixture was diluted with dichloromethane and washed with water. Theorganic phases were dried (Na₂SO₄), concentrated and purified on silicagel (Biotage Isolera 1, 4 g column, EtOAc/hexanes 0-50-100%, producteluted at 50-100%, two peaks) to provide a clear glass. An off-whitesolid was obtained by slow evaporation of a MeOH/water solution. ¹HNMR(400 MHz, CHLOROFORM-d) δ ppm 8.43-8.58 (m, 1H), 8.05 (s, 1H), 6.38 (dd,J=7.39, 4.77 Hz, 1H), 4.78 (q, J=6.76 Hz, 1H), 4.47 (d, J=1.91 Hz, 2H),2.90-3.09 (m, 3H), 2.82 (s, 1H), 2.71 (dt, J=13.71, 7.33 Hz, 1H), 2.45(d, J=6.44 Hz, 1H), 2.36 (td, J=7.57, 2.50 Hz, 2H), 1.72-1.88 (m, 2H),1.54-1.69 (m, 2H), 1.21-1.51 (m, 18H), 0.81-0.99 (m, 6H). LCMS(M−1)=558.4.

Example 43:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methylheptanoate

Step 1:((2R,3S)-5-(6-amino-2-fluoro-9H-purin-9-v)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylheptanoate. To a solution of(2R,3S)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol(500 mg, 1.705 mmol) in acetonitrile (70 mL) {used sonicator todissolve} was added heptanoic acid (0.254 mL, 1.790 mmol) and2-chloro-1-methylpyridin-1-ium iodide (566 mg, 2.216 mmol) and themixture was stirred at ambient temperature for 15 min. The mixture wascooled to −15° C. Ice/EtOH) and triethylamine (0.713 mL, 5.11 mmol) wasadded followed by N,N-dimethylpyridin-4-amine (20.83 mg, 0.170 mmol).After 2 h 45 min (final temperature −5° C.), 0.5 M HCl/water (100 mL)was added and the mixture was extracted with dichloromethane. Theorganic phases were washed with NaHCO₃/water, dried (Na₂SO₄),concentrated and purified on silica gel (Biotage Isolera 1, 80 g Goldcolumn, MeOH/dichloromethane 3% isocratic) to provide((2R,3S)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylheptanoate (330 mg, 0.814 mmol, 47.7% yield) (N70935-59-1, fractions52-73) as an off-white solid. ¹HNMR (400 MHz, DMSO-d6) δ ppm 8.26 (s,1H), 7.81 (br s, 2H), 6.25 (dd, J=8.11, 4.05 Hz, 1H), 5.76 (d, J=5.25Hz, 1H), 4.63-4.79 (m, 1H), 4.42 (d, J=11.68 Hz, 1H), 4.10 (d, J=11.68Hz, 1H), 3.61 (s, 1H), 2.80 (ddd, J=13.35, 7.27, 3.93 Hz, 1H), 2.5-2.4(m, 1H), 2.05-2.31 (m, 2H), 1.34-1.55 (m, 2H), 1.08-1.29 (m, 6H), 0.83(t, J=7.03 Hz, 3H). LCMS (M+1)=406.2.

Step 2:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)ox)tetrahydrofuran-2-yl)methyl heptanoate. To a solution of((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylheptanoate (320 mg, 0.789 mmol) in DCM (40 mL) was added Et3N (0.440 mL,3.16 mmol) and the mixture was cooled to 0° C. then hexylcarbonochloridate (0.258 mL, 1.579 mmol) was added dropwise. After 1 hat 0° C. the mixture was stirred at ambient temperature for 1 h. DMAP(9.64 mg, 0.079 mmol) was added and after 3.5 h at ambient temperaturethe mixture was diluted with dichloromethane and washed with saturatedNaHCO₃/water. The organic phase was dried (Na₂SO₄), concentrated andpurified on silica gel (Biotage Isolera 1, 24 g column, EtOAc/hexanes0-50-100%) to provide((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy)tetrahydro-furan-2-yl)methylheptanoate (132 mg, 0.247 mmol, 31.3% yield) as a white solid (afterconcentration from MeCN). ¹HNMR (400 MHz, CHLOROFORM-d) δ ppm 7.78-8.02(m, 1H), 6.39 (t, J=6.44 Hz, 1H), 5.76 (br s, 2H), 5.61 (dd, J=7.27,5.60 Hz, 1H), 4.56 (d, J=12.16 Hz, 1H), 4.42 (d, J=12.16 Hz, 1H), 4.22(td, J=6.74, 2.98 Hz, 2H), 3.09-3.23 (m, 1H), 2.76-2.88 (m, 1H), 2.71(s, 1H), 2.37 (td, J=7.57, 4.41 Hz, 2H), 1.68-1.80 (m, 2H), 1.60-1.68(m, 2H), 1.21-1.48 (m, 12H), 0.91 (dt, J=12.28, 6.97 Hz, 6H). LCMS(M+1)=534.6.

Example 44:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methyldecanoate

Step 1:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl hexyl carbonate. To a stirredsolution of(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-2-ethynyltetrahydrofuran-3-ol(500 mg, 0.940 mmol), DMAP (115 mg, 0.940 mmol) and triethylamine (0.393mL, 2.82 mmol) at 0° C. was slowly added a solution of hexylcarbonochloridate (0.169 mL, 1.034 mmol) in DCM (1.33 mL) and themixture was stirred at 0° C. for ˜ 2 hours. Sat'd NaHCO₃ was added andthe mixture was extracted with EtOAc. The extracts were washed withbrine, dried over Na₂SO₄, filtered and concentrated. The residue waspurified on silica gel (40 g gold column, 0-70% hexanes/EtOAc) to affordtitle compound as white residue (293 mg, 47%). LCMS (M+1)=660.4.

Step 2:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetra-hydrofuran-3-ylhexyl carbonate. To a solution of(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-2-ethynyltetrahydrofuran-3-ylhexyl carbonate (347 mg, 0.526 mmol) in THF (5.0 mL) at ambienttemperature was added TBAF (0.789 mL, 0.789 mmol) and the mixture wasstirred for 30 minutes. The mixture was concentrated and then purifiedon silica gel (0-10% DCM/MeOH) to afford title compound as white solid(179 mg, 81%). LCMS (M+1)=422.2.

Step 3:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)ox)tetrahydrofuran-2-yl)methyl decanoate. To a mixture of(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ylhexyl carbonate (179 mg, 0.425 mmol), decanoic acid (80 mg, 0.467 mmol),DMAP (51.9 mg, 0.425 mmol) and DIEA (0.371 mL, 2.124 mmol) was added EDC(163 mg, 0.849 mmol) at ambient temperature and the mixture was allowedto stir overnight. The mixture was concentrated and then purified onsilica gel (0-10% DCM/MeOH) to afford title compound as white solid (173mg, 71%). ¹HNMR (400 MHz, DMSO-d6) b=8.31 (s, 1H), 7.85 (br s, 2H),6.38-6.32 (m, 1H), 5.59 (dd, J=5.4, 7.0 Hz, 1H), 4.43 (d, J=11.7 Hz,1H), 4.25 (d, J=11.7 Hz, 1H), 4.19-4.11 (m, 2H), 3.79 (s, 1H), 3.25-3.13(m, 1H), 2.77-2.61 (m, 1H), 2.38-2.12 (m, 2H), 1.71-1.57 (m, 2H),1.51-1.41 (m, 2H), 1.39-1.13 (m, 18H), 0.92-0.81 (m, 6H); LCMS(M+1)=576.4.

Example 45:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methylbenzoate

Step 1:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methylbenzoate. A suspension of((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methanol(920 mg, 2.258 mmol) in DCM (22 mL) was treated with benzoic acid (331mg, 2.71 mmol), DMAP (276 mg, 2.258 mmol), EDC (779 mg, 4.06 mmol), DIEA(1.971 mL, 11.29 mmol), and stirred at rt for 18 hours. The reaction wasdiluted with water, extracted with DCM, washed with sat. NaHCO₃, brine,dried over Na₂SO₄, filtered, and concentrated. Purification with columnchromatography (0-100% EtOAc/DCM) afforded((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methylbenzoate (984.4 mg, 1.924 mmol, 85% yield) as white solid. ¹HNMR (400MHz, CHLOROFORM-d) δ ppm 7.97-7.90 (m, 2H), 7.81 (s, 1H), 7.58-7.51 (m,1H), 7.45-7.34 (m, 2H), 6.25 (dd, J=3.5, 8.0 Hz, 1H), 5.66 (br s, 2H),5.19 (dd, J=7.0, 8.2 Hz, 1H), 4.74 (d, J=12.2 Hz, 1H), 4.44 (d, J=12.2Hz, 1H), 3.03 (ddd, J=3.3, 7.1, 13.2 Hz, 1H), 2.74-2.62 (m, 2H), 0.96(s, 9H), 0.18 (s, 6H); LCMS (m/z) ES⁺=512.2 (M+1), ES⁻=510.4 (M−1).

Step 2:((2R,3S,5R)-3-((tert-butyldimethylsily)oxy)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)tetrahydrofuran-2-yl)methylbenzoate. A solution of((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methylbenzoate (475 mg, 0.928 mmol) in DCM (9.2 mL) was treated with Et3N(0.388 mL, 2.79 mmol), pyridine (0.225 mL, 2.79 mmol), tetradecanoylchloride (0.379 mL, 1.393 mmol), DMAP (11.34 mg, 0.093 mmol), and thenstirred at rt for 18 hours. The reaction was diluted with water,extracted with DCM, washed with aq. sat. NaHCO₃, brine, dried overNa₂SO₄, filtered, and concentrated. Purification with columnchromatography (0-100% EtOAc/Hexane) afforded((2R,3S,5R)-3-((tert-butyldimethyl-silyl)oxy)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)tetrahydrofuran-2-yl)methylbenzoate (209.8 mg, 0.291 mmol, 31.3% yield) as yellow film. ¹HNMR (400MHz, CHLOROFORM-d) δ ppm 8.33 (s, 1H), 7.96 (s, 1H), 7.92-7.86 (m, 2H),7.58-7.49 (m, 1H), 7.44-7.35 (m, 2H), 6.28 (dd, J=3.5, 8.0 Hz, 1H), 5.15(dd, J=6.9, 8.1 Hz, 1H), 4.77 (d, J=12.2 Hz, 1H), 4.43 (d, J=12.2 Hz,1H), 3.03 (ddd, J=3.3, 7.0, 13.3 Hz, 1H), 2.93 (t, J=7.5 Hz, 2H),2.78-2.64 (m, 2H), 1.77 (quin, J=7.5 Hz, 2H), 1.50-1.39 (m, 2H),1.39-1.19 (m, 18H), 0.97 (s, 9H), 0.92-0.84 (m, 3H), 0.29-0.10 (m, 6H);LCMS (m/z) ES⁺=722.5 (M+1), ES⁻=720.6 (M−1).

Step 3:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methylbenzoate. An ice cold solution of((2R,3S,5R)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)tetrahydrofuran-2-yl)methylbenzoate (285 mg, 0.395 mmol) in THF (8 mL) was treated with TBAF (1 Min THF) (0.592 mL, 0.592 mmol), and stirred at 0° C. for 2 hours. Themixture was treated with additional 1M TBAF (150 uL) and stirred at 0°C. for 40 min. The reaction was quenched with AcOH (˜0.5 mL), dilutedwith water, and extracted with EtOAc. The combined organics was washedwith brine, dried over Na₂SO₄, filtered, and concentrated. Purificationwith column chromatography [0-65% (3:1 EtOAc:EtOH)/Hexane] gave((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methylbenzoate (194.4 mg, 0.310 mmol, 79% yield) as off-white solid. ¹HNMR(400 MHz, CHLOROFORM-d) δ ppm 8.39 (s, 1H), 8.02-7.93 (m, 3H), 7.62-7.54(m, 1H), 7.48-7.39 (m, 2H), 6.37 (dd, J=4.5, 7.6 Hz, 1H), 4.95 (q, J=7.0Hz, 1H), 4.79-4.61 (m, 2H), 3.08 (ddd, J=4.5, 6.8, 13.7 Hz, 1H), 2.94(t, J=7.4 Hz, 2H), 2.85 (s, 1H), 2.73 (td, J=7.5, 13.6 Hz, 1H), 2.48 (d,J=6.7 Hz, 1H), 1.76 (quin, J=7.5 Hz, 2H), 1.49-1.39 (m, 2H), 1.38-1.20(m, 18H), 0.96-0.83 (m, 3H); LCMS (m/z) ES⁺=608.4 (M+1), ES⁻=606.4(M−1).

Example 46:((2R,3S,5R)-5-(6-decanamido-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylbenzoate

Step 1:((2R,3S,5R)-3-((tert-butyldimethylsilyl)oxy)-5-(6-decanamido-2-fluoro-9H-purin-9-vi)-2-ethynyltetrahydrofuran-2-yl)methylbenzoate. A solution of((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methylbenzoate (470 mg, 0.919 mmol) in DCM (9.2 mL) was treated with Et3N(0.384 mL, 2.76 mmol), pyridine (0.223 mL, 2.76 mmol), decanoyl chloride(0.286 mL, 1.378 mmol), and DMAP (11.22 mg, 0.092 mmol). The reactionwas stirred at rt for 18 hours. The reaction was diluted with water,extracted with DCM, washed with aq. sat. NaHCO₃, brine, dried overNa₂SO₄, filtered, and concentrated. Purification with columnchromatography (0-100% EtOAc/Hexane) gave((2R,3S,5R)-3-((tert-butyldimethylsilyl)oxy)-5-(6-decanamido-2-fluoro-9H-purin-9-yl)-2-ethynyltetrahydrofuran-2-yl)methylbenzoate (217.7 mg, 0.327 mmol, 35.6% yield) as yellow film. NMRcontained aliphatic impurity. ¹HNMR (400 MHz, CHLOROFORM-d) δ ppm 8.32(s, 1H), 7.96 (s, 1H), 7.93-7.86 (m, 2H), 7.57-7.49 (m, 1H), 7.44-7.36(m, 2H), 6.28 (dd, J=3.6, 7.9 Hz, 1H), 5.15 (dd, J=7.0, 8.2 Hz, 1H),4.77 (d, J=12.2 Hz, 1H), 4.43 (d, J=12.2 Hz, 1H), 3.03 (ddd, J=3.5, 7.0,13.2 Hz, 1H), 2.97-2.88 (m, 2H), 2.78-2.63 (m, 2H), 1.77 (quin, J=7.5Hz, 2H), 1.48-1.20 (m, 12H), 0.97 (s, 9H), 0.92-0.84 (m, 3H), 0.21-0.16(m, 6H); LCMS (m/z) ES⁺=666.5 (M+1), ES⁻=664.6 (M−1).

Step 2:((2R,3S,5R)-5-(6-decanamido-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl)methylbenzoate. An ice cold solution of((2R,3S,5R)-3-((tert-butyldimethylsilyl)oxy)-5-(6-decanamido-2-fluoro-9H-purin-9-yl)-2-ethynyltetrahydrofuran-2-yl)methylbenzoate (272 mg, 0.408 mmol) in THF (8 mL) was treated with TBAF (1M inTHF) (0.613 mL, 0.613 mmol), and stirred at 0° C. for 2 hours. Themixture was treated with additional 1M TBAF (100 uL) and stirred at 0°C. for 40 min. The reaction was quenched with AcOH (˜0.5 mL), dilutedwith water, and extracted with EtOAc. The combined organics was washedwith brine, dried over Na₂SO₄, filtered, and concentrated. Purificationwith column chromatography [0-65% (3:1 EtOAc:EtOH)/Hexane] gave((2R,3S,5R)-5-(6-decanamido-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylbenzoate (164.1 mg, 0.292 mmol, 71.4% yield) as light yellow solid.¹HNMR (400 MHz, CHLOROFORM-d) δ ppm 8.35 (s, 1H), 8.04-7.93 (m, 3H),7.62-7.54 (m, 1H), 7.47-7.37 (m, 2H), 6.37 (dd, J=4.5, 7.6 Hz, 1H), 4.95(q, J=7.1 Hz, 1H), 4.78-4.62 (m, 2H), 3.08 (ddd, J=4.5, 6.9, 13.6 Hz,1H), 2.94 (t, J=7.4 Hz, 2H), 2.85 (s, 1H), 2.73 (td, J=7.4, 13.6 Hz,1H), 2.45 (d, J=6.9 Hz, 1H), 1.77 (quin, J=7.5 Hz, 2H), 1.47-1.38 (m,2H), 1.38-1.20 (m, 10H), 0.94-0.85 (m, 3H); LCMS (m/z) ES⁻=550.4 (M−1).

Example 47:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methylheptanoate

Step 1:((2R,3S,5R)-3-((tert-butyldimethylsily)oxy)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)tetrahydrofuran-2-yl)methylheptanoate. To a solution of((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methylheptanoate (280 mg, 0.539 mmol) and DIEA (0.565 mL, 3.23 mmol) in DCM (7mL) was added DMAP (32.9 mg, 0.269 mmol) followed by dropwise additionof a solution of tetradecanoyl chloride (266 mg, 1.078 mmol) indichloromethane (0.5 mL). After 2.5 h more tetradecanoyl chloride (0.09mL) was added. After 2 h saturated NaHCO₃/water was added and theorganic phases were dried (Na₂SO₄), concentrated, and purified on silicagel (Biotage Isolera 1, 12 g column, 0-100% EtOAc/hexanes 0-100%) toprovide((2R,3S,5R)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)tetrahydrofuran-2-yl)methylheptanoate (153 mg, 0.207 mmol, 38.5% yield) as a yellowish oil whichwas in the next step.

Step 2:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methylheptanoate. To a solution of((2R,3S,5R)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)tetra-hydrofuran-2-yl)methylheptanoate (150 mg, 0.205 mmol) in THF (2 mL) at 0° C. was added TBAF(0.226 mL, 0.226 mmol) (1 M/THF) and the mixture was stirred at 0° C.for 3 h (10° C. final bath temperature). acetic acid (0.013 mL, 0.226mmol) was added and the mixture was concentrated. The residue waspurified on silica gel (Biotage Isolera 1, 4 g column, EtOAc/hexanes0-100%, product eluted at 90-100%) to provide((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methylheptanoate (109.7 mg, 0.176 mmol, 86% yield) as an off-white solid (bydissolving in a small amount of MeOH, adding 2 drops of water and slowevaporation in open container, dried in vacuo). ¹HNMR (400 MHz,CHLOROFORM-d) δ ppm 8.51-8.64 (m, 1H), 8.06 (s, 1H), 6.38 (br dd,J=6.91, 4.77 Hz, 1H), 4.78 (br d, J=6.68 Hz, 1H), 4.47 (s, 2H),2.91-3.11 (m, 4H), 2.82 (s, 1H), 2.62-2.78 (m, 1H), 2.49 (br d, J=6.44Hz, 1H), 2.26-2.43 (m, 2H), 1.71-1.92 (m, 3H), 1.14-1.54 (m, 26H),0.82-1.02 (m, 6H). LCMS (M+1)=617.5.

Example 48:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methyldecanoate

Step 1:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methyldecanoate. To a mixture of((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl) diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetra-hydrofuran-2-yl)methanol(3 g, 3.58 mmol), Et3N (2.495 mL, 17.90 mmol) and DMAP (0.656 g, 5.37mmol) in DCM (40 mL) was added Et3N (2.495 mL, 17.90 mmol). The reactionmixture was stirred at r.t. for 2 h. LCMS indicated completion ofreaction. The reaction mixture was quenched with water (50 mL),extracted with DCM (30 mL*3), the organic phases were combined, washedwith brine (30 mL), dried over Na₂SO₄, concentrated under vacuum. Theresidue was purified by flash chromatography (silica gel, 120 g, pet.ether:EtOAc=4:1) to give((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenyl-methoxy)tetrahydrofuran-2-yl)methyldecanoate (2.7 g, 1.788 mmol, 49.9% yield)) as a yellow solid. LCMS(ESI) m/z calcd for C₆₂H₆₂FN₅O₆: 991 Found: 992 (M+H)⁺. ¹HNMR (400 MHz,CDCl3) δ 8.21 (s, 1H), 7.57-7.50 (m, 4H), 7.40-7.18 (m, 20H), 7.04 (s,1H), 6.81-6.75 (m, 4H), 6.08 (dd, J=3.2, 8.0 Hz, 1H), 4.31 (d, J=12.0Hz, 1H), 4.05 (d, J=12.0 Hz, 1H), 3.78-3.72 (m, 6H), 3.00-2.91 (m, 1H),2.45-2.01 (m, 4H), 1.67-1.59 (m, 2H), 1.33-1.18 (m, 12H), 0.87 (t, J=7.2Hz, 3H).

Step 2:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydro-furan-2-yl)methyldecanoate. To a solution((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenyl-methoxy)tetrahydrofuran-2-yl)methyl decanoate (2.7 g, 2.72 mmol) in DCM (30 mL)stirred at 15° C. was added TFA (5 mL, 64.9 mmol) dropwise. The reactionmixture was stirred at 15° C. for 1 h. LCMS indicated completion ofreaction. The reaction mixture was quenched with NaHCO₃ (30 ml), andextracted with DCM (30 ml×2), the organic phases were combined, washedwith brine (15 ml), dried over Na₂SO₄, concentrated under vacuum. Theresidue was purified by flash chromatography (silica gel, 120 g,DCM:MeOH=10:1) to give((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydro-furan-2-yl)methyldecanoate (1 g, 2.201 mmol, 81% yield) as a white solid. LCMS (ESI) m/zcalcd for C₂₂H₃₀FN₅O₄: 447 Found: 448 (M+H)⁺. ¹HNMR (400 MHz, CDCl3) δ7.88 (s, 1H), 6.31 (dd, J=4.4, 7.6 Hz, 1H), 6.10 (br, s, 2H), 4.79 (t,J=7.2 Hz, 1H), 4.50-4.40 (m, 2H), 2.99-2.92 (m, 1H), 2.81-2.60 (m, 2H),2.38-2.30 (m, 2H), 1.65-1.51 (m, 2H), 1.34-1.14 (m, 12H), 0.91-0.75 (m,3H).

Step 3:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methyldecanoate. To a solution of((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyldecanoate (1 g, 2.235 mmol) in DCM (60 mL) was added imidazole (1.521 g,22.35 mmol) and tert-butylchlorodimethylsilane (1.347 g, 8.94 mmol), andthe resulting mixture was stirred at 30° C. for overnight. LCMSindicated completion of reaction. The reaction mixture was quenched withwater (50 mL), extracted with DCM (20 mL*3), the organic phases werecombined, washed with brine (50 mL), dried over Na₂SO₄, concentratedunder vacuum. The residue was purified by gel silica column (120 g,EtOAc:pet. Ether=1:1) to give((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methyldecanoate (1.2 g, 98.09%, yield: 94%) as a white solid. LCMS (ESI) m/zcalcd for C₂₈H₄₄FN₅O₄Si: 561. Found: 562 (M+H)⁺. ¹HNMR (400 MHz, CDCl3)δ 7.86 (s, 1H), 6.25 (dd, J=4.0, 8.0 Hz, 1H), 5.94 (br, s, 2H), 4.93 (t,J=7.6 Hz, 1H), 4.45 (d, J=12.0 Hz, 1H), 4.23 (d, J=12.0 Hz, 1H),2.89-2.82 (m, 1H), 2.68-2.58 (m, 2H), 2.30-2.19 (m, 2H), 1.61-1.50 (m,2H), 1.33-1.17 (m, 12H), 0.93 (s, 9H), 0.87 (t, J=7.2 Hz, 3H), 0.13 (d,J=4.8 Hz, 6H).

Step 4:((2R,3S,5R)-3-((tert-butyldimethylsily)oxy)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)tetrahydrofuran-2-yl)methyldecanoate. To a solution of((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methyldecanoate (1.2 g, 2.136 mmol) in DCM (10 mL) was added DMAP (0.391 g,3.20 mmol), Et3N (1.191 mL, 8.54 mmol) and tetradecanoyl chloride (1.318g, 5.34 mmol). The reaction mixture was stirred at r.t. for 4 h. LCMSindicated completion of reaction. The reaction mixture was quenched withwater (10 mL) and extracted with DCM (10 mL×3), the organic phases werecombined, washed with brine (10 mL), dried over Na₂SO₄, concentratedunder vacuum. The residue was purified by gel silica column (80 g,EtOAc:pet. ether=4:1) to give((2R,3S,5R)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)tetrahydro-furan-2-yl)methyldecanoate (700 mg, 85%, 19.52% yield) as a white semisolid. LCMS (ESI)m/z calcd for Ca₂H₇₀FN₅O₅Si: 771. Found: 772 (M+H)⁺. ¹HNMR (400 MHz,CDCl3) δ 8.88 (s, 1H), 8.08 (d, J=1.2 Hz, 1H), 6.30 (dd, J=3.6, 7.6 Hz,1H), 4.89 (t, J=7.2 Hz, 1H), 4.44 (d, J=12.0 Hz, 1H), 4.24 (d, J=12.0Hz, 1H), 2.96-2.80 (m, 3H), 2.71-2.60 (m, 2H), 2.28-2.22 (m, 2H),1.80-1.68 (m, 2H), 1.60-1.50 (m, 2H), 1.37-1.17 (m, 32H), 0.93 (s, 9H),0.90-0.81 (m, 6H), 0.13 (d, J=5.2 Hz, 6H).

Step 5:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methyldecanoate. To a stirred solution of((2R,3S,5R)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)tetrahydrofuran-2-yl)methyldecanoate (700 mg, 0.907 mmol) in THF (5 mL) was added TBAF (1.08 ml, 1mol/L in THF). The resulting mixture was stirred for 2 hour at roomtemperature. LCMS indicated completion of reaction. The majority ofsolvent was removed under vacuum. The residue was purified by gel silicacolumn (80 g, EtOAc: pet. ether=4:1) to give the desired product(purity: 96%) as a white solid. The solid was triturated with ACN (30ml) for 16 h. The resulting solid was filtered through a Buchner funnel,rinsed with ACN, dried beside sunlight lamp (T=50° C.) for 6 h, and wasstored in a cool dry place overnight to give((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methyldecanoate (151 mg, 99.05%, yield: 25.08%) as a white solid. LCMS (ESI)m/z calcd for C₃₆H₅₆FN₅O₅: 657. Found: 658 (M+1)⁺. ¹HNMR (400 MHz,CDCl3) δ 8.66 (s, 1H), 8.06 (s, 1H), 6.36 (dd, J=4.4, 7.6 Hz, 1H), 4.77(t, J=6.4 Hz, 1H), 4.49-4.41 (m, 2H), 2.98-2.81 (m, 3H), 2.74 (s, 1H),2.72-2.68 (m, 1H), 2.36-2.32 (m, 2H), 1.79-1.56 (m, 6H), 1.44-1.20 (m,30H), 0.89-0.85 (m, 6H).

Example 49:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methyl2-propylpentanoate

Step 1:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methyl2-propylpentanoate. To a stirred solution of((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methanol(200 mg, 0.491 mmol), EDC (141 mg, 0.736 mmol), and DMAP (60.0 mg, 0.491mmol)) in DCM (6 mL) was added 2-propylpentanoic acid (0.079 mL, 0.491mmol) followed by DIEA (0.214 mL, 1.227 mmol) and the mixture wasstirred at r.t. for 1 h. The reaction mixture was concentrated andpurified by Isco, 40 gram gold column (DCM/MeOH 0-10%) to give thedesired product as a white solid (238 mg, 85%). LCMS (M+1)=535.3.

Step 2:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methyl2-propylpentanoate. To a solution of((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methyl2-propylpentanoate (238 mg, 0.446 mmol), DMAP (5.45 mg, 0.045 mmol) and2-chloro-1-methylpyridinium iodide (285 mg, 1.115 mmol) in acetonitrile(6.0 mL) and THF (6.0 mL) was added tetradecanoic acid (255 mg, 1.115mmol) followed by Et3N (0.373 mL, 2.68 mmol) and the mixture was stirredat 60° C. in a sealed tube overnight. The mixture was diluted with DCMand washed with sat. aq. NaHCO₃, the organic layer dried over sodiumsulfate and concentrated. Purification by prep. TLC (Hexanes/EtOAc 1:1)afforded the desired product as a solid. ¹HNMR (400 MHz, CHLOROFORM-d) δppm 0.16 (s, 3H) 0.17 (s, 3H) 0.80-0.92 (m, 9H) 0.96 (s, 9H) 1.17-1.47(m, 24H) 1.48-1.56 (m, 2H) 1.62-1.71 (m, 2H) 1.71-1.81 (m, 2H) 2.32(ddd, J=8.70, 5.36, 3.34 Hz, 1H) 2.35-2.46 (m, 1H) 2.63 (s, 1H) 2.70(dt, J=13.41, 7.72 Hz, 1H) 2.85-2.95 (m, 2H) 4.26 (d, J=12.16 Hz, 1H)4.44 (d, J=12.16 Hz, 1H) 4.89 (t, J=7.27 Hz, 1H) 6.34 (dd, J=7.63, 3.81Hz, 1H) 8.13 (s, 1H) 9.38 (br s, 1H).

To a solution of above solid in THF (5.00 mL) and added TBAF (1M, THF)(0.491 mL, 0.491 mmol) at 0° C. and the mixture was stirred at 0° C. for1 h and then added acetic acid (0.028 mL, 0.491 mmol) at 0° C., allowedto warm to r.t and concentrated. Purification by Isco, 40 gram goldcolumn (DCM/MeOH 0-5%) afforded the desired product as a solid (98 mg,34%). ¹HNMR (400 MHz, CHLOROFORM-d) δ ppm 0.82-0.92 (m, 9H) 1.22-1.48(m, 26H) 1.51-1.66 (m, 2H) 1.74-1.90 (m, 2H) 2.36-2.48 (m, 1H) 2.73 (dt,J=13.71, 6.97 Hz, 1H) 2.82 (s, 1H) 2.92-3.05 (m, 3H) 4.41-4.49 (m, 2H)4.76 (t, J=6.68 Hz, 1H) 6.41 (dd, J=7.15, 4.77 Hz, 1H) 8.12 (s, 1H) 8.82(s, 1H). LCMS (M+1)=630.5. LCMS (M−1)=628.5.

Example 50:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl3-(2-acetoxy-4,6-dimethylphenyl)-3-methylbutanoate

Step 1:9-((2R,4S,5R)-5-(((tert-butyldiphenylsilyl)oxy)methyl)-5-ethynyl-4-((4-methoxyphen-yl)diphenylmethoxy)tetrahydrofuran-2-yl)-2-fluoro-N-((4-methoxy-phenyl)diphenylmethyl)-9H-purin-6-amine.To a stirred suspension of(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-2-ethynyltetra-hydrofuran-3-ol(2.66 g, 5.00 mmol) in DCM (60 mL) was added silver nitrate (2.55 g,15.01 mmol), 2,4,6-trimethylpyridine (4.00 mL, 30.0 mmol) and4-methoxytriphenylmethyl chloride (4.63 g, 15.01 mmol). The resultingsuspension was stirred at ambient temperature for 2 hours. The mixturewas diluted with EtOAc and filtered over Celite. The filtrate was washedwith 10% citric acid (2×), followed by sat'd NaHCO₃ (2×), dried overNa₂SO₄, filtered and concentrated. The residue was purified on silicagel (120 g column, 0-100% hexanes/EtOAc) to afford title compound aswhite solid (5.138 g, 95%). LCMS (M+1)=1076.57.

Step 2:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methanol.To stirred solution of9-((2R,4S,5R)-5-(((tert-butyldiphenylsilyl)oxy)methyl)-5-ethynyl-4-((4-methoxyphen-yl)diphenylmethoxy)tetrahydrofuran-2-yl)-2-fluoro-N-((4-methoxyphenyl)diphenylmethyl)-9H-purin-6-amine (5.138 g, 4.77 mmol) in THF (45 mL) wasadded 1M TBAF/THF (7.16 mL, 7.16 mmol) and the mixture was allowed tostir at ambient temperature for 90 minutes. The mixture was concentratedand the residue was purified on silica gel (120 g column, 0-100%hexanes/EtOAc) to afford title compound as white solid (3.908 g, 98%).LCMS (M+1)=838.42.

Step 3:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methyl3-(2-acetoxy-4,6-dimethylphenyl)-3-methylbutanoate. To a mixture of((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxy-phenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methanol (315 mg, 0.376 mmol),3-(2-acetoxy-4,6-dimethylphenyl)-3-methylbutanoic acid (119 mg, 0.451mmol), DMAP (45.9 mg, 0.376 mmol) and DIEA (0.328 mL, 1.880 mmol) wasadded EDC (144 mg, 0.752 mmol) at ambient temperature and the mixturewas allowed to stir overnight. The mixture was concentrated and thenpurified on silica gel (24 g column, 0-70% hexanes/EtOAc) to affordtitle compound as colorless residue (366 mg, 90%). LCMS (M+1)=1084.61;LCMS (M−1)=1083.79.

Step 4:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydro-furan-2-yl)methyl3-(2-acetoxy-4,6-dimethylphenyl)-3-methylbutanoate. A solution of((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxy-phenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methyl3-(2-acetoxy-4,6-dimethylphenyl)-3-methylbutanoate (366 mg, 0.338 mmol)in DCM (1.6 mL) was treated with formic Acid (0.4 mL) and triethylsilane(0.108 mL, 0.675 mmol) and then stirred at ambient temperature for 3hours. The mixture was concentrated and then purified on silica gel (40g column, 0-10% DCM/MeOH) to afford title compound as white solid (118mg, 65%). ¹HNMR (400 MHz, DMSO-d6) δ=8.25 (s, 1H), 7.82 (br s, 2H),6.77-6.71 (m, 1H), 6.56-6.53 (m, 1H), 6.24 (dd, J=4.3, 7.9 Hz, 1H), 5.71(d, J=5.5 Hz, 1H), 4.66-4.59 (m, 1H), 4.27 (d, J=11.7 Hz, 1H), 4.00 (d,J=11.9 Hz, 1H), 3.59 (s, 1H), 2.84-2.71 (m, 2H), 2.61 (d, J=16.0 Hz,1H), 2.53-2.43 (m, 1H, overlapping DMSO peak), 2.41 (s, 3H), 2.18 (s,3H), 2.14 (s, 3H), 1.40 (s, 3H), 1.35 (s, 3H). LCMS (M+1)=540.31.

Example 51:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((heptanoyloxy)methyl)tetrahydrofuran-3-yltetradecanoate

Step 1:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methylheptanoate. To a mixture of((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydro-furan-2-yl)methanol(420 mg, 0.50 mmol) in DCM (10 mL) was added DMAP (61.2 mg, 0.50 mmol),Et3N (0.28 mL, 2.00 mmol) and heptanoyl chloride (149 mg, 1.00 mmol).The reaction mixture was stirred for 16 hours. LCMS indicated completionof reaction. The reaction mixture was diluted with water (10 mL),extracted with DCM (10 mL×3). The combined organic phases were washedwith water (20 mL), brine (10 mL) and dried over Na₂SO₄. Afterfiltration, the filtrate was concentrated to dryness under vacuum. Theresidue was purified by silica gel column (120 g, pet. ether:EtOAc=1:1)to afford((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methylheptanoate (300 mg, 63% yield) as a pale-yellow solid. LCMS (ESI) m/zcalcd for C₅₉H₅₆FN₅O₆:949. Found: 950 (M+H)⁺. ¹HNMR (300 MHz, DMSO-d₆) δ8.06 (s, 1H), 7.96 (s, 1H), 7.53-7.51 (m, 4H), 7.40-7.17 (m, 20H),6.92-6.78 (m, 4H), 6.13 (dd, J=7.8, 3.9 Hz, 1H), 4.68 (t, J=7.2 Hz, 1H),4.18 (d, J=12.0 Hz, 1H), 3.97 (s, 1H), 3.79 (d, J=12.0 Hz, 1H), 3.72 (s,3H), 3.68 (s, 3H), 2.09-2.00 (m, 1H), 1.97-1.79 (m, 3H), 1.37-1.01 (m,8H), 0.80 (t, J=6.9 Hz, 3H).

Step 2:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-v)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylheptanoate.To the mixture of((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenyl-methoxy)tetrahydrofuran-2-yl)methylheptanoate (300 mg, 0.32 mmol) in DCM (6 mL) was added TFA (1 mL)dropwise at 0° C. The reaction mixture was stirred for 2 h at 0° C. LCMSindicated completion of reaction. The reaction mixture was concentratedto dryness under vacuum. The residue was re-dissolved in DCM (10 mL) andwashed with NaHCO₃ (aq, 10 mL), brine (10 mL), dried over Na₂SO₄. Afterfiltration, the filtrate was concentrated to dryness under vacuum toafford((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylheptanoate (120 mg, yield: 93%) as a white solid. The crude product wasused to next step without further purification. LCMS (ESI) m/z calcd forC19H24FN5O4:405, Found: 406 (M+H)⁺.

Step 3:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((heptanoyloxy)methyl)tetra-hydrofuran-3-yl tetradecanoate. The mixture of tetradecanoic acid(214 mg, 0.94 mmol), EDCl (241 mg, 1.26 mmol) and DMAP (153 mg, 1.25mmol) in DMF (4 mL) was stirred for 2 h at 25° C. To the above mixturewas added((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylheptanoate (120 mg, 0.30 mmol). The reaction mixture was stirred at 25°C. for 16 h. LCMS indicated completion of reaction. The reaction mixturewas quenched with water (10 mL) and extracted with EtOAc (10 mL×3). Thecombine organic phases were washed with brine (10 mL) and dried overNa₂SO₄. After filtration, the solvent was removed under vacuum. Theresidue was purified by Prep-HPLC: Column: XSelect CSH Prep C18 OBDColumn, 5 um, 19×150 mm; Mobile Phase A: water (0.05% TFA), Mobile PhaseB: ACN; Flow rate: 25 mL/min; Gradient: 90 B to 95 B in 8 min; 254 nm;Rt: 6.48 min. The collected fraction (RT: 6.48 min) was lyophilized toafford(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((heptanoyloxy)methyl)tetrahydro-furan-3-yltetradecanoate (93.7 mg, purity: 99.66%, yield: 51%) as white solid.LCMS (ESI) m/z calcd for C₃₃H₅₀FN₅O₅:615. Found: 616 (M+H)⁺. ¹HNMR (300MHz, DMSO-d₆) δ 8.33 (s, 1H), 7.90 (br s, 2H), 6.34 (t, J=6.6 Hz, 1H),5.70 (t, J=5.4 Hz, 1H), 4.40 (d, J=11.4 Hz, 1H), 4.22 (d, J=11.4 Hz,1H), 3.79 (s, 1H), 3.17-3.13 (m, 1H), 2.66-2.57 (m, 1H), 2.42-2.37 (m,2H), 2.32-2.21 (m, 2H), 1.60-1.40 (m, 4H), 1.39-1.22 (m, 26H), 0.89-0.84(m, 6H).

Example 52:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((tridecyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methylheptanoate

Step 1:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methylheptanoate. To a mixture of((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydro-furan-2-yl)methanol(1.0 g, 1.19 mmol) in DCM (10 mL) was added DMAP (0.15 g, 1.19 mmol),Et3N (0.67 mL, 4.77 mmol) and heptanoyl chloride (0.355 g, 2.387 mmol).The reaction mixture was stirred for 16 hours. LCMS indicated completionof reaction. The reaction mixture was diluted with water (10 mL),extracted with DCM (10 mL×3). The combined organic phases were washedwith water (20 mL), brine (10 mL) and dried over Na₂SO₄. Afterfiltration, the filtrate was concentrated to dryness under vacuum. Theresidue was purified by silica gel column (120 g, pet. ether:EtOAc=1:1)to afford((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenyl-methoxy)tetrahydrofuran-2-yl)methylheptanoate (800 mg, yield: 70.6%) as a white solid. LCMS (ESI) m/z calcdfor C₅₉H₅₆FN₅O₆: 949. Found: 950 (M+H)⁺. ¹HNMR (300 MHz, DMSO-d₆) δ 8.06(s, 1H), 7.94 (s, 1H), 7.52-7.49 (m, 4H), 7.43-7.12 (m, 20H), 6.96-6.76(m, 4H), 6.13 (dd, J=8.0, 4.0 Hz, 1H), 4.68 (t, J=7.6 Hz, 1H), 4.19 (d,J=12.0 Hz, 1H), 3.96 (s, 1H), 3.80 (d, J=12.0 Hz, 1H), 3.72 (s, 3H),3.69 (s, 3H), 2.09-2.00 (m, 1H), 1.96-1.80 (m, 3H), 1.41-0.95 (m, 8H),0.81-0.78 (m, 3H).

Step 2:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydro-furan-2-yl)methylheptanoate. To the mixture of((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenyl-methoxy)tetrahydrofuran-2-yl)methyl heptanoate (800 mg, 0.84 mmol) in DCM (20mL) was added TFA (2.0 mL) dropwise at 0° C. The reaction mixture wasstirred for 2 h at 0° C. LCMS indicated completion of reaction. Thereaction mixture was concentrated to dryness under vacuum. The residuewas dissolved in DCM (20 mL) and washed with NaHCO₃ (aq, 10 mL), brine(10 mL), dried over Na₂SO₄. After filtration, the filtrate wasconcentrated to dryness under vacuum. The crude product was purified byreverse phase Column: C18 (80 g); Mobile Phase A: water (1% FA), MobilePhase B: ACN; Flow rate: 60 mL/min; Gradient: 0% B to 100% B in 30 min;254/220 nm. The collected fractions were concentrated to afford((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylheptanoate (247 mg, yield: 72.4%) as white solid. LCMS (ESI) m/z calcdfor C₁₉H₂₄FN₅O₄: 405 Found: 406 (M+H)⁺.

Step 3:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((tridecyloxy)carbonyl) oxy) tetrahydrofuran-2-yl)methyl heptanoate. To a stirred mixture of((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylheptanoate (218 mg, 0.54 mmol) and K₂CO₃ (220 mg, 1.59 mmol) in THF (5mL) was added di(1H-imidazol-1-yl)methanone (150 mg, 0.93 mmol) at 25°C. The reaction mixture was stirred at 25° C. for 2 h. To the abovemixture was added Cs2CO3 (380 mg, 1.17 mmol) and tridecan-1-ol (550 mg,2.74 mmol). The reaction mixture was stirred at 25° C. for 1 h. LCMSindicated completion of reaction. The reaction mixture was quenched withwater (10 mL) and extracted with EtOAc (10 mL×3). The organic phaseswere washed with brine (20 mL), dried over Na₂SO₄, concentrated underreduced pressure. The crude product was purified by PreP-HPLC: Column:XBridge Prep OBD C18 Column, 30×150 mm 5 um; Mobile Phase A: water(0.05% TFA), Mobile Phase B: ACN; Flow rate: 254 mL/min; Gradient: 55 Bto 72 B in 7 min; 254 nm; Rt: 6.52 min. The collected fraction (RT: 6.52min) was lyophilized to afford((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((tridecyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methylheptanoate (150 mg, purity: 98.7%, yield: 44%) as white solid. LCMS(ESI) m/z calcd for C₃₃H₅₀FN₅O₆: 631. Found: 632 (M+H)⁺. ¹HNMR (400 MHz,DMSO-d₆) δ 8.33 (s, 1H), 8.07-7.72 (m, 2H), 6.35 (t, J=6.8 Hz, 1H), 5.59(dd, J=6.8, 5.2 Hz, 1H), 4.42 (d, J=11.6 Hz, 1H), 4.23 (d, J=11.6 Hz,1H), 4.19-4.07 (m, 2H), 3.81 (s, 1H), 3.25-3.11 (m, 1H), 2.73-2.61 (m,1H), 2.35-2.16 (m, 2H), 1.68-1.56 (m, 2H), 1.50-1.38 (m, 2H), 1.30-1.15(m, 26H), 0.90-0.79 (m, 6H).

Example 53:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((propionyloxy)methyl)tetrahydrofuran-3-ylstearate

Step 1:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methylpropionate. To a stirred solution of((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxy-phenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methanol (700 mg, 0.84 mmol), Et3N (0.70 mL, 5.01mmol) and DMAP (51 mg, 0.42 mmol) in DCM (7 mL) was added propionylchloride (232 mg, 2.506 mmol) dropwise at 0° C. The reaction mixture wasstirred at 0° C. for 30 m. LCMS indicated completion of reaction. Thereaction was quenched by the addition of water (3 mL), the organiclayers were separated and the water phase was extracted with DCM (2mL*3). The combined organic layers were dried over Na₂SO₄ and evaporatedto dryness under vacuum to give the crude title compound (800 mg, 94.5%,yield: 89%) as yellow oil. LCMS (ESI) m/z calcd for C₅₅H₄₃FN₅O₆: 893.Found: 894 (M+1)⁺. ¹HNMR (400 MHz, DMSO-d₆) δ 8.07 (s, 1H), 7.99 (s,1H), 7.60-7.45 (m, 4H), 7.40-7.14 (m, 20H), 6.91-6.77 (m, 4H), 6.15-6.12(m, 1H), 4.72 (t, J=7.6 Hz, 1H), 4.18 (d, J=12.0 Hz, 1H), 3.96 (s, 1H),3.75 (d, J=12.0 Hz, 1H), 3.69 (s, 3H), 3.67 (s, 3H), 2.13-2.02 (m, 1H),1.94-1.90 (m, 3H), 0.75 (t, J=7.6 Hz, 3H).

Step 2:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydro-furan-2-yl)methylpropionate. A solution of((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)di-phenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenyl-methoxy)tetrahydrofuran-2-yl)methylpropionate (630 mg, 0.71 mmol) in DCM (7 mL) was added trifluoroaceticacid (TFA) (0.050 mL). The reaction solution was stirred at 25° C. for30 min. LCMS indicated completion of reaction. The reaction was dilutedwith methanol until the yellow solution turn to colorless andconcentrated under vacuum. The residue was subjected to HPLCpurification (C18, 0-80% ACN/water with 0.1% FA) to give the desiredproduct (200 mg, 98.7%, yield: 80%) as white solid. LCMS (ESI) m/z calcdfor C₁₅H₁₆FN₅O₄: 349. Found: 350 (M+1)⁺. ¹HNMR (400 MHz, DMSO-d₆) δ 8.28(s, 1H), 7.86 (br s, 2H), 6.25 (dd, J=8.0, 4.0 Hz, 1H), 5.79 (d, J=5.6Hz, 1H), 4.74-4.68 (m, 1H), 4.42 (d, J=11.6 Hz, 1H), 4.11 (d, J=11.6 Hz,1H), 3.64 (s, 1H), 2.90-2.76 (m, 1H), 2.49-2.44 (m, 1H), 2.35-2.15 (m,2H), 0.95 (t, J=7.6 Hz, 3H).

Step 3:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((propionyloxy)methyl)tetrahydrofuran-3-yl stearate. A solution of stearic acid (489 mg, 1.72mmol) in DMF (8 mL) was added EDC (439 mg, 2.29 mmol) and DMAP (280 mg,2.29 mmol). The reaction mixture was stirred at 25° C. for 30 min. Then((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylpropionate (200 mg, 0.57 mmol) was added. The resulting mixture wasstirred at 25° C. for 20 hours. LCMS indicated completion of reaction.The reaction was diluted with water (30 ml), and the resulting mixturewas extracted with EtOAc (20 ml*3). The combined organic layers werewashed with brine (20 mL), dried over anhydrous sodium sulfate andevaporated to dryness under vacuum. The residue was purified byPrep-HPLC (Column: XSelect CSH Prep C18 OBD Column, 5 um, 19*150 mm;Mobile Phase A: Water (0.05% TFA), Mobile Phase B: ACN; Flow rate: 25mL/min; Gradient: 95 B to 95 B in 7 min; 254 nm, RT: 6.05 min)) to givethe desired product (156.8 g, 99.12%, yield: 44%) as white solid. LCMS(ESI) m/z calcd for C₃₃H₅₀FN₅O₅: 615. Found: 616 (M+1)⁺. ¹HNMR (400 MHz,DMSO-d₆) δ 8.34 (s, 1H), 7.95 (brs, 2H), 6.34 (t, J=6.8 Hz, 1H), 5.70(t, J=5.6 Hz, 1H), 4.42 (d, J=11.6 Hz, 1H), 4.21 (d, J=11.6 Hz, 1H),3.79 (s, 1H), 3.15-3.12 (m, 1H), 2.62-2.58 (m, 1H), 2.41-2.25 (m, 4H),1.60-1.56 (m, 2H), 1.25-1.23 (m, 28H), 0.98 (t, J=7.6 Hz, 3H), 0.85 (t,J=6.4 Hz, 3H).

Example 54:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-heptanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methyl2-propylpentanoate

Step 1:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxypheny)diphenylmethoxy)tetrahydrofuran-2-yl)methyl 2-propylpentanoate. To astirred solution of((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxy-phenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenyl-methoxy)tetrahydro-furan-2-yl)methanol(2.00 g, 2.387 mmol), Et3N (1.0 mL, 7.16 mmol) and DMAP (146 mg, 1.193mmol) in DCM (20.00 mL) was added a solution of 2-propylpentanoylchloride (427 mg, 2.63 mmol) in DCM (2 mL) dropwise over 5 min at 0° C.The reaction mixture was stirred at 20° C. for 2 hr. LCMS indicatedcompletion of reaction. The reaction mixture was quenched with water,extracted with EtOAc (3×20 mL). The organic phases were combined, washedwith brine, dried over Na₂SO₄ and concentrated under vacuum. The residuewas subjected to Prep-TLC (EtOAc:pet. ether=1:1) to give the desiredproduct (1.7 g, 99%, yield: 73%) as off white solid. LCMS (ESI) m/zcalcd for C₆₀H₅₈FN₅O₆: 963. Found: 964 (M+1)⁺. ¹HNMR (400 MHz,Chloroform-d) δ 7.61 (s, 1H), 7.59-7.54 (m, 4H), 7.44-7.21 (m, 20H),7.10 (br s, 1H), 6.89-6.78 (m, 4H), 6.14-6.10 (m, 1H), 4.61 (t, J=7.6Hz, 1H), 4.35 (d, J=12.0 Hz, 1H), 4.19 (d, J=12.0 Hz, 1H), 3.80 (s, 3H),3.78 (s, 3H), 2.85 (s, 1H), 2.24-2.03 (m, 2H), 1.73-1.60 (m, 2H),1.46-1.32 (m, 3H), 1.17-1.04 (m, 4H), 0.82-0.78 (m, 6H).

Step 2:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl2-propylpentanoate. To the stirred solution of((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methyl 2-propylpentanoate (1700 mg, 1.763 mmol) inDCM (20 mL) was added trifluoroacetic acid (TFA) (2 mL). The resultingmixture was stirred for 1 hour at room temperature. LCMS indicatedcompletion of reaction. The reaction was quenched by pouring into sodiumcarbonate (aq.) and the mixture was extracted with EtOAc (3×20 mL). Theorganic phases were combined, washed with saturated brine, dried overNa₂SO₄ and concentrated under vacuum. The residue was purified by flashchromatography (silica gel, 120 g, EtOAc:pet. ether=1:1) to give thedesired product as white solid (650 mg, 98%, yield: 86%). LCMS (ESI) m/zcalcd for C₂₀H₂₆FN₅O₄: 419. Found: 420 (M+1)⁺. ¹HNMR (400 MHz,Chloroform-d) b 7.91 (s, 1H), 6.39-6.34 (m, 1H), 6.12 (br s, 2H), 4.77(t, J=6.8 Hz, 1H), 4.46 (d, J=1.2 Hz, 2H), 3.04-2.98 (m, 1H), 2.80 (s,1H), 2.73-2.66 (m, 2H), 2.46-2.39 (m, 1H), 1.66-1.52 (m, 2H), 1.50-1.37(m, 2H), 1.35-1.18 (m, 4H), 0.89-0.85 (m, 6H).

Step 3:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methyl2-propylpentanoate. To a solution of((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl2-propylpenta-noate (640 mg, 1.526 mmol) and imidazole (312 mg, 4.58mmol) in DMF (10.00 mL) was added a solution of TBS-Cl (690 mg, 4.58mmol) in DMF (1 mL). The reaction mixture was stirred at 40° C. for 16hr. LCMS indicated completion of reaction. The reaction was quenched bypouring into brine and the mixture was extracted with EtOAc (3×20 mL).The organic phases were combined, washed with saturated brine, driedover Na₂SO₄ and concentrated under vacuum. The residue was purified byflash chromatography (silica gel, 40 g, EtOAc:pet. ether=1:1) to givethe desired product (500 mg, 58%, yield: 95%) as off white solid. LCMS(ESI) m/z calcd for C₂₆H₄₀FN₅O₄Si: 533. Found: 534 (M+1)⁺. ¹HNMR (400MHz, Chloroform-d) δ 7.91 (s, 1H), 6.29 (dd, J=7.6, 3.6 Hz, 1H), 5.93(br s, 2H), 4.94 (t, J=7.2 Hz, 1H), 4.46 (d, J=12.0 Hz, 1H), 4.24 (d,J=12.0 Hz, 1H), 2.96-2.91 (m, 1H), 2.70-2.63 (m, 1H), 2.61 (s, 1H),2.36-2.29 (m, 1H), 1.61-1.44 (m, 2H), 1.44-1.31 (m, 2H), 1.31-1.18 (m,4H), 0.96 (s, 9H), 0.87-0.82 (m, 6H), 0.17 (s, 6H).

Step 4:((2R,3S,5R)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyl-5-(2-fluoro-6-heptanamido-9H-purin-9-yl)tetrahydrofuran-2-yl)methyl2-propylpentanoate.((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methyl2-propylpenta-noate (750 mg, 1.405 mmol) was dissolved in DCM (20 mL).Et3N (0.979 mL, 7.03 mmol) and DMAP (86 mg, 0.703 mmol) were added, andthen heptanoyl chloride (418 mg, 2.81 mmol) was added at 0° C. Thereaction mixture was stirred at 0° C. for overnight. LCMS indicatedcompletion of reaction. The solvent was removed under vacuum. Theresidue was subjected to Prep-TLC (pet. ether:EtOAc=1:1) to give thedesired product (400 mg, 92%, yield: 42%) as off white oil. LCMS (ESI)m/z calcd for C₃₃H₅₂FN₅O₅Si: 645. Found: 646 (M+1)⁺. ¹HNMR (400 MHz,Chloroform-d) δ 8.70 (s, 1H), 8.09 (s, 1H), 6.34 (dd, J=7.6, 4.0 Hz,1H), 4.91 (t, J=7.2 Hz, 1H), 4.45 (d, J=12.0 Hz, 1H), 4.26 (d, J=12.0Hz, 1H), 3.00-2.87 (m, 3H), 2.73-2.65 (m, 1H), 2.63 (s, 1H), 2.37-2.29(m, 1H), 1.82-1.74 (m, 2H), 1.61-1.10 (m, 14H), 0.96 (s, 9H), 0.88-0.79(m, 9H). 0.17 (s, 6H).

Step 5:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-heptanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methyl2-propylpentanoate. To the stirred solution of((2R,3S,5R)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyl-5-(2-fluoro-6-heptanamido-9H-purin-9-yl)tetrahydrofuran-2-yl)methyl2-propylpentanoate (400 mg, 0.619 mmol) in tetrahydrofuran (THF) (10 mL)was added TBAF (1.2 mL, 1M in THF, 1.2 mmol). The resulting mixture wasstirred for 1 hour at room temperature. LCMS indicated completion ofreaction. The reaction mixture was concentrated under vacuum. Water wasadded, the mixture was extracted with EtOAc (3×20 mL). The combinedorganic phases were washed with brine, dried over Na₂SO₄ andconcentrated under vacuum. The residue was purified by Prep-TLC(MeOH:DCM=1:10) to give crude product. The crude product was re-purifiedby Prep-HPLC (Column: XSelect CSH Prep C18 OBD Column, 5 um, 19*150 mm;Mobile Phase A: Water (0.05% TFA), Mobile Phase B: ACN; Flow rate: 25mL/min; Gradient: 65 B to 86 B in 6 min; 254 nm; RT: 4.07) The collectedfraction was lyophilized to give the desired product (150 mg, 97%,yield: 44%), LCMS (ESI) m/z calcd for C₂₇H₃₈FN₅O₅: 531. Found: 532(M+1)⁺. ¹HNMR (400 MHz, Chloroform-d) δ 8.69 (s, 1H), 8.08 (s, 1H), 6.41(dd, J=7.6, 5.2 Hz, 1H), 4.75 (t, J=6.8 Hz, 1H), 4.44-4.43 (m, 2H),3.00-2.92 (m, 3H), 2.80 (s, 1H), 2.74-2.69 (m, 1H), 2.42-2.38 (m, 1H),2.15 (br s, 1H), 1.78-1.72 (m, 2H), 1.60-1.52 (m, 2H), 1.46-1.22 (m,12H), 0.91-0.83 (m, 9H).

Example 55:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methylpropionate

Step 1:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methylpropionate. To a stirred mixture of((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxy-phenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methanol (2.5 g, 2.98 mmol), Et3N (1.25 mL, 8.94mmol) and DMAP (226 mg, 1.85 mmol) in DCM (30 mL) was added propionylchloride (550 mg, 5.94 mmol) at 0° C. The reaction mixture was stirredat 0° C. for 30 min. LCMS indicated completion of reaction. The reactionmixture was quenched with water (50 mL) and extracted with DCM (30×3mL). The combined organic layers were washed with brine (30 mL) anddried over Na₂SO₄. After filtration, the filtrate was concentrated todryness under vacuum. The crude product was purified by silica gelcolumn (120 g, EtOAc:pet. ether=1:1) to afford((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methylpropionate (2.18 g, yield: 73.6%) as a white solid. LCMS (ESI) m/z calcdfor C₅₅H₄₈FN₅O₆: 893. Found: 894 (M+H)⁺. ¹HNMR (300 MHz, DMSO-d₆) δ 8.07(s, 1H), 7.99 (s, 1H), 7.58-7.43 (m, 4H), 7.40-7.14 (m, 20H), 6.86-6.81(m, 4H), 6.16-6.12 (m, 1H), 4.74-4.69 (m, 1H), 4.18 (d, J=12.0 Hz, 1H),3.96 (s, 1H), 3.74 (d, J=12.0 Hz, 1H), 3.72 (s, 3H), 3.69 (s, 3H),2.13-2.07 (m, 1H), 1.97-1.82 (m, 3H), 0.75 (t, J=7.5 Hz, 3H).

Step 2:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydro-furan-2-yl)methylpropionate. To a stirred mixture of((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methyl propionate (2.18 g, 2.44 mmol) in DCM (50mL) was added TFA (5.4 mL) at 0° C. The reaction mixture was stirred at0° C. for 2 h. LCMS indicated completion of reaction. The reactionmixture was quenched with water (20 mL) and was basified to pH=8 withsat.NaHCO₃.(aq). The precipitated solids were collected by filtrationand washed with water to afford((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylpropionate (660 mg, yield: 62%) as white solid. The crude product wasused in the next step directly without further purification. LCMS (ESI)m/z calcd for C₁₅H₁₆FN₅O₄: 349 Found: 350 (M+H)⁺.

Step 3:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methylpropionate. To a stirred mixture of((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methylpropionate (640 mg, 1.83 mmol) and imidazole (1.3 g, 19.1 mmol) in DMF(10 mL) was added TBS-Cl (1.1 g, 7.30 mmol) at 25° C. The reactionmixture was stirred at 30° C. for 12 h. LCMS indicated completion ofreaction. The reaction mixture was quenched with water (20 mL),extracted with EtOAc (3×20 mL). The combined organic layers were washedwith brine (20 mL), dried over with Na₂SO₄, filtered and concentratedunder reduced pressure. The crude product was purified by silica gel (80g, pet. ether/EtOAc=1:1) to afford((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methylpropionate (730 mg, yield: 84%) as a white solid. LCMS (ESI) m/z calcdfor C₂₁H₃₀FN₅O₄Si: 463. Found: 464 (M+H)⁺. ¹HNMR (400 MHz, DMSO-d₆) δ8.29 (s, 1H), 7.89 (br s, 2H), 6.30-6.27 (m, 1H), 4.97 (t, J=6.8 Hz,1H), 4.34 (d, J=12.0 Hz, 1H), 4.12 (d, J=12.0 Hz, 1H), 3.67 (s, 1H),3.09-2.85 (m, 1H), 2.50-2.37 (m, 1H), 2.31-2.01 (m, 2H), 0.98-0.90 (m,12H), 0.15 (s, 6H).

Step 4:((2R,3S,5R)-3-((tert-butyldimethylsily)oxy)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)tetrahydrofuran-2-yl)methylpropionate. To a stirred mixture of((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methylpropionate (580 mg, 1.25 mmol), Et3N (1.38 mL, 9.88 mmol) and DMAP (153mg, 1.25 mmol) in DCM (10 mL) was added tetradecanoyl chloride (1.23 g,4.98 mmol) at 0° C. The reaction mixture was stirred at 25° C. for 12 h.LCMS indicated completion of reaction. The reaction mixture was quenchedwith water (20 mL), extracted with EtOAc (3×20 mL). The combined organiclayers were dried over with Na₂SO₄, filtered and concentrated underreduced pressure. The crude product was purified by silica gel column(80 g, pet. ether/EtOAc=1:1) to afford((2R,3S,5R)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)tetrahydrofuran-2-yl)methylpropionate (490 mg, yield: 52.3%) as a yellow oil. LCMS (ESI) m/z calcdfor C₃₅H₅₆FN₅O₅Si: 673. Found: 674 (M+H)⁺. ¹HNMR (300 MHz, DMSO-d₆) δ11.02 (s, 1H), 8.60 (s, 1H), 6.41-6.37 (m, 1H), 4.97 (t, J=9.0 Hz, 1H),4.35 (d, J=12.0 Hz, 1H), 4.15 (d, J=12.0 Hz, 1H), 3.69 (s, 1H),3.09-2.88 (m, 1H), 2.58 (d, J=7.2 Hz, 2H), 2.26-2.13 (m, 2H), 1.66-1.50(m, 2H), 1.34-1.17 (m, 24H), 0.96-0.83 (m, 12H), 0.15 (s, 6H).

Step 5:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methylpropionate. To a stirred mixture of((2R,3S,5R)-3-((tert-butyldimethylsilyl)oxy)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)tetrahydrofuran-2-yl)methylpropionate (470 mg, 0.70 mmol) in THF (20 mL) was added TBAF (1.15 mL,1.15 mmol, 1N in THF) at 0° C. The reaction mixture was stirred at 0° C.for 30 min. LCMS indicated completion of reaction. The reaction mixturewas quenched with water (20 mL), extracted with EtOAc (3×20 mL). Thecombined organic layers were washed with sat.NH₄Cl (aq., 3×20 mL), driedover with Na₂SO₄, filtered and concentrated under reduced pressure. Thecrude product was purified by Prep-HPLC: Column: XSelect CSH Prep C18OBD Column, 5 um, 19×150 mm; Mobile Phase A: water (0.1% FA), MobilePhase B: ACN; Flow rate: 25 mL/min; Gradient: 67 B to 95 B in 8 min; 254nm; Rt: 7.23 min. The collected fraction (RT: 7.23 min) was lyophilizedto afford((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-tetradecanamido-9H-purin-9-yl)-3-hydroxytetra-hydrofuran-2-yl)methylpropionate (252 mg, purity: 99.04%, yield: 64%) as a white solid. LCMS(ESI) m/z calcd for C₂₉H₄₂FN₅O₅ 559. Found: 560 (M+H)⁺. ¹HNMR (400 MHz,DMSO-d₆) δ 11.0 (s, 1H), 8.59 (s, 1H), 6.36-633 (m, 1H), 5.84 (d, J=5.2Hz, 1H), 4.73-4.71 (m, 1H), 4.42 (d, J=11.6 Hz, 1H), 4.13 (d, J=11.6 Hz,1H), 3.66 (s, 1H), 2.90-2.78 (m, 1H), 2.57-2.51 (m, 3H), 2.25-2.19 (m,2H), 1.60-1.57 (m, 2H), 1.28-1.23 (m, 20H), 0.93 (t, J=7.6 Hz, 3H), 0.85(t, J=6.4 Hz, 3H).

Example 56:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((ethoxycarbonyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methyltetradecanoate

Step 1:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methyltetradecanoate. To a stirred solution of((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenyl-methoxy)tetrahydrofuran-2-yl)methanol(550 mg, 0.656 mmol), Et3N (0.27 mL, 1.97 mmol) and DMAP (40 mg, 0.328mmol) in DCM (5 mL) was added tetradecanoyl chloride (486 mg, 1.969mmol) at 0° C. The reaction mixture was stirred at 0° C. for 30 min.LCMS indicated completion of reaction. The reaction was quenched withwater (3 mL). The organic layers were separated out. The water phase wasextracted with DCM (2 mL*3). The combined organic layers were dried overanhydrous sodium sulfate and evaporated to dryness concentrated undervacuum. The residue was purified by Prep-TLC (pet. ether:EtOAc=1:1) togive the desired product (638 mg, 92%, yield: 85%) as a yellow solid.LCMS (ESI) m/z calcd for C₆₆H₇₀FN₅O₆: 1048. Found: 1049 (M+1)⁺. ¹HNMR(300 MHz, DMSO-d₆) δ 8.06 (s, 1H), 7.92 (s, 1H), 7.56-7.46 (m, 4H),7.42-7.13 (m, 20H), 6.86-6.83 (m, 4H), 6.15-6.11 (m, 1H), 4.71-4.65 (m,1H), 4.18 (d, J=12.0 Hz, 1H), 3.96 (s, 1H), 3.79 (d, J=12.0 Hz, 1H),3.72 (s, 3H), 3.68 (s, 3H), 1.93-1.85 (m, 2H), 1.64 (br s, 2H),1.24-1.15 (m, 22H), 0.86-0.82 (m, 3H).

Step 2:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyltetradecanoate. A stirred solution of((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)di-phenylmethoxy)tetrahydrofuran-2-yl)methyl tetradecanoate (638 mg, 0.61 mmol) in DCM (6mL) was added trifluoroacetic acid (TFA) (0.6 mL). The reaction mixturewas stirred at 25° C. for 30 minutes in air. LCMS indicated completionof reaction. The reaction mixture was quenched by the addition ofmethanol until the red solution turn to colorless and concentrated undervacuum. The residue was purified by Prep-HPLC purification (C18, 0-100%ACN/water with 0.1% FA) to give the desired product (270 mg, 100%,yield: 88%). LCMS (ESI) m/z calcd for C₂₆H₃₈FN₅O₄: 503. Found: 504(M+1)⁺. ¹HNMR (300 MHz, DMSO-d₆) δ 8.27 (s, 1H), 7.85 (br s, 2H),6.26-6.22 (m, 1H), 5.79 (d, J=5.4 Hz, 1H), 4.74-4.67 (m, 1H), 4.42 (d,J=12.0 Hz, 1H), 4.10 (d, J=12.0 Hz, 1H), 3.63 (s, 1H), 2.86-2.70 (m,1H), 2.47-2.46 (m, 1H), 2.30-2.10 (m, 2H), 1.41 (br s, 2H), 1.41-1.17(m, 20H), 0.90-0.80 (m, 3H).

Step 3:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((ethoxycarbonyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methyltetradecanoate. To a stirred solution of((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyltetradecanoate (270 mg, 0.536 mmol) and K₂CO₃ (222 mg, 1.608 mmol) inTHF (2 mL) was added solid CDI (261 mg, 1.608 mmol) at 25° C. Thereaction mixture was stirred at 25° C. for 30 min. LCMS indicatedcompletion of reaction. The reaction was quenched with ice cooled-water(8 mL). The resulting mixture was extracted with EtOAc (10 ml*3). Thecombined organic layers were washed with brine (10 mL), dried overanhydrous sodium sulfate and evaporated to dryness in vacuum to give thedesired product (250 mg, 95%, yield: 74.1%) as white solid. LCMS (ESI)m/z calcd for C₃₀H₄₀FN₇O₅: 597. Found: 598 (M+1)⁺.

Step 4:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-((ethoxycarbonyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methyltetradecanoate. To a stirred solution of(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((tetradecanoyloxy)methyl)tetrahydrofuran-3-yl1H-imidazole-1-carboxylate (250 mg, 0.418 mmol) and Cs2CO3 (291 mg,0.894 mmol) in tetrahydrofuran (THF) (4 mL) was added ethanol (193 mg,4.18 mmol) at 25° C. The reaction mixture was stirred at 25° C. for 2hours. LCMS indicated completion of reaction. The reaction was quenchedwith ice cooled-water (8 mL). The resulting mixture was extracted withEtOAc (10 ml*3). The combined organic layers were washed with brine (10mL), dried over anhydrous sodium sulfate and evaporated to dryness undervacuum. The residue was purified by Prep-HPLC (Column: XBridge Prep C18OBD Column, 19*150 mm 5 um; Mobile Phase A: Water (0.1% FA), MobilePhase B: ACN; Flow rate: 25 mL/min; Gradient: 75 B to 95 B in 7 min; 220nm, 254 nm. RT: 6.06 min) to give the desired product (158 mg, 98.84%,yield: 64.4%) as a white solid. LCMS (ESI) m/z calcd for C₂₉H₄₂FN₅O₆:615. Found: 616 (M+1)⁺. ¹HNMR (400 MHz, DMSO-d₆) δ 8.32 (s, 1H), 7.89(br s, 2H), 6.34 (t, J=6.8 Hz, 1H), 5.58 (t, J=5.2 Hz, 1H), 4.42 (d,J=11.2 Hz, 1H), 4.25-4.17 (m, 3H), 3.82 (s, 1H), 3.23-3.16 (m, 1H),2.70-2.64 (m, 1H), 2.33-2.18 (m, 2H), 1.43-1.42 (m, 2H), 1.27-1.19 (m,23H), 0.85 (t, J=6.8 Hz, 3H).

Example 57:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl(1,3-bis(isobutyryloxy)propan-2-yl) succinate

Step 1: 2-oxopropane-1,3-diyl bis(2-methylpropanoate). To a solution of1,3-dihydroxypropan-2-one (30 g, 333 mmol) in DCM (200 mL) stirred undernitrogen at 0° C. was added DMAP (1.22 g, 9.99 mmol), pyridine (81 mL,999 mmol) and isobutyryl chloride (78 g, 733 mmol) in DCM (100 mL)dropwise during 30 min. The reaction mixture was stirred at 0° C. forovernight. TLC showed the reaction was completed (pet. ether:EtOAc=1:1,colored by KMnO₄). The reaction mixture was quenched with saturatedNaHCO₃ aq. (100 ml) and the organic layers were washed with brine (100mL*6). The combined organic layers were dried over Na₂SO₄ andconcentrated under vacuum. The residue was purified by flashchromatography (silica gel, 330 g, pet. ether:EtOAc=5:1) to give thedesired product (34 g, 90%, yield: 44.3%) as a yellow oil. ¹HNMR (400MHz, Chloroform-d) b 4.77 (s, 4H), 2.60-2.81 (m, 2H), 1.18-1.25 (m,12H).

Step 2: 2-hydroxypropane-1,3-diyl bis(2-methylpropanoate). To a solutionof 2-oxopropane-1,3-diyl bis(2-methylpropanoate) (10 g, 43.4 mmol) inTHF (100 mL)/water (10 mL) stirred at 0° C. was added NaBH₄ (1.97 g,52.1 mmol) portionwise. The reaction mixture was stirred at 0° C. for 30min. TLC traces showed the reaction was completed (pet. ether:EtOAc=1:3,colored by KMnO₄). The reaction mixture was quenched with HCl (1 mmol/L,100 mL), extracted with EtOAc (80 mL*3). The combined organic layerswere dried over Na₂SO₄ and concentrated under vacuum to give the desiredproduct (9 g, 90%, yield: 80%) as a yellow oil. ¹HNMR (400 MHz,Chloroform-d) δ 3.81-4.73 (m, 4H), 2.56-2.71 (m, 2H), 1.18-1.25 (m,12H).

Step 3: 4-((1,3-bis(isobutyryloxy)propan-2-yl)oxy)-4-oxobutanoic acid.2-Hydroxypropane-1,3-diyl bis(2-methylpropanoate) (10 g, 43.1 mmol) wasdissolved in dichloromethane (DCM) (30 mL)/THF (30 mL)/Pyridine (30 mL),dihydrofuran-2,5-dione (8.62 g, 86 mmol) and DMAP (0.53 g, 4.31 mmol)were added, the resulting mixture was stirred for 6.5 hours at 60° C.LCMS indicated completion of reaction. The reaction mixture was quenchedwith HCl (1 M, 80 mL), extracted with EtOAc (80 mL*3). The combinedorganic layers were dried over Na₂SO₄ and concentrated under vacuum. Theresidue was purified by flash chromatography (silica gel, 330 g, pet.ether:EtOAc=3:2) to give the title compound (13 g, 95%, yield: 86%) as acolorless oil. LCMS (ESI) m/z calcd for C₁₅H₂₄O₈: 332 Found: 355(M+Na)⁺. ¹HNMR (400 MHz, Chloroform-d) δ5.27-5.36 (m, 1H), 3.96-4.49 (m,4H), 2.44-2.80 (m, 6H), 1.17-1.21 (m, 12H).

Step 4: 1,3-bis(isobutyryloxy)propan-2-yl(((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenyl-methoxy)tetrahydrofuran-2-yl)methyl)succinate. 4-((1,3-bis(isobutyryloxy)propan-2-yl)oxy)-4-oxobutanoic acid(397 mg, 1.19 mmol) was dissolved in DMF (20 mL), DMAP (365 mg, 2.99mmol) and 3-(((ethylimino)methyl-ene)amino)-N,N-dimethylpropan-1-aminehydrochloride (572 mg, 2.99 mmol) were added, the resulting mixture wasstirred for 2 hours at 20° C. Then,((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenyl-methyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenyl-methoxy)tetrahydrofuran-2-yl)methanol(500 mg, 0.60 mmol) was added and the resulting mixture was stirred forovernight at 20° C. LCMS indicated completion of reaction. The reactionwas quenched with water (80 mL) and the mixture was extracted with EtOAc(50 mL*3). The combined organic layers were dried over Na₂SO₄ andconcentrated under vacuum. The residue was purified by flashchromatography (silica gel, 120 g, pet. ether:EtOAc=1:1) to give thedesired product (600 mg, 95%, yield: 83%) as a white solid. LCMS (ESI)m/z calcd for C₆₇H₆₆FN₅O₁₂: 1151. Found: 1152 (M+H)⁺. ¹HNMR (400 MHz,Chloroform-d) δ 7.62 (s, 1H), 7.54-7.57 (m, 4H), 7.38-7.46 (m, 2H),7.23-7.36 (m, 18H), 7.07 (s, 1H), 6.76-6.88 (m, 4H), 6.11 (dd, J=8.0,3.6 Hz, 1H), 5.22-5.35 (m, 1H), 4.64 (t, J=8.0 Hz, 1H), 4.26-4.41 (m,3H), 4.15-4.20 (m, 2H), 3.96-4.06 (m, 1H), 3.79 (d, J=16.4 Hz, 6H), 2.86(s, 1H), 2.15-2.69 (m, 8H), 1.17-1.25 (m, 12H).

Step 5:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydro-furan-2-yl)methyl(1,3-bis(isobutyryloxy)propan-2-yl) succinate. To a solution of1,3-bis(isobutyryloxy) propan-2-yl(((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxy-phenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methyl) succinate (600 mg, 0.52 mmol) in DCM (10mL) stirred at 15° C. was added TFA (1 ml) dropwise. The reactionmixture was stirred at 15° C. for 1 hour. LCMS indicated completion ofreaction. The reaction mixture was quenched with saturated NaHCO₃ aq (30ml), and extracted with DCM (15 ml*2). The combined organic phases werewashed with brine (15 ml) and concentrated under vacuum. The residue waspurified by Prep-HPLC purification (Column: XBridge Prep OBDC18 Column,19*250 mm, 5 um; Mobile Phase A: Water (10 MMOL/L NH₄HCO₃+0.1% NH₃.H₂O),Mobile Phase B: ACN; Flow rate: 25 mL/min; Gradient: 35 B to 57 B in 11min; 254 nm; RT: 9.13 min). The collected fraction (RT: 9.13 min) waslyophilized to give the desired product (103 mg, 98.2%, yield: 32%) as awhite solid. LCMS (ESI) m/z calcd for C₂₇H₃₄FN₅O₁₀: 607. Found: 608(M+H)⁺. ¹HNMR (400 MHz, DMSO-d₆) δ 8.27 (s, 1H), 7.85 (br, s, 2H), 6.25(dd, J=8.0, 4.4 Hz, 1H), 5.80 (d, J=5.2, 1H), 5.17 (dd, J=6.4, 4.0 Hz,1H), 4.68 (dd, J=13.2, 7.6 Hz, 1H), 4.39 (d, J=11.6, 1H), 4.26-4.09 (m,5H), 3.62 (s, 1H), 2.80-2.73 (m, 1H), 2.54-2.47 (m, 7H), 1.05 (d, J=6.4Hz, 12H).

Example 58:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((dodecanoyloxy)methyl)-2-ethynyltetrahydrofuran-3-yldodecanoate

To a stirred mixture of(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol(500 mg, 1.705 mmol), Et3N (0.951 mL, 6.82 mmol) and DMAP (41.7 mg,0.341 mmol) in DCM (30 mL) was added dodecanoyl chloride (783 mg, 3.58mmol) dropwise. The reaction mixture was stirred at room temperature for2 h. LCMS indicated completion of reaction. The reaction was quenchedwith water (50 ml), and extracted with DCM (3×50 ml). The combinedorganic phases were dried over sodium sulphate and concentrated underreduced pressure to afford crude product as a yellow solid. The samplewas further purified on a silica column (120 g) using 0%-80% EtOAc/pet.ether solvent gradient over 80 mins, Flow rate: 70 mL/min. Theappropriate fractions containing desired product were identified by UVabsorbance (254 nm), combined and evaporated in vacuo to give whitesolid. The solid was triturated with ACN (15 ml), stirred at roomtemperature overnight. The resulting solid was filtered through aBuchner funnel, rinsed with ACN, dried under sun lamp for 1 h, and wasput in a cool dry place overnight. Then the solid was collected to give(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((dodecanoyloxy)methyl)-2-ethynyltetrahydrofuran-3-yldodecanoate (520 mg, 0.783 mmol, 45.9% yield) as a white solid. LCMS(ESI) m/z calcd for C₃₆H₅₆FN₅O₅: 657. Found: 658 (M+H)⁺. ¹HNMR (400 MHz,Chloroform-d) δ 7.95 (s, 1H), 6.40 (t, J=6.4 Hz, 1H), 6.09 (br s, 2H),5.65 (dd, J=7.2, 5.6 Hz, 1H), 4.49 (d, J=12.0 Hz, 1H), 4.39 (d, J=12.0Hz, 1H), 2.95-3.02 (m, 1H), 2.66-2.75 (m, 2H), 2.30-2.43 (m, 4H),1.57-1.69 (m, 4H), 1.17-1.40 (m, 32H), 0.85-0.90 (m, 6H).

Example 59:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((palmitoyloxy)methyl)tetrahydrofuran-3-ylpalmitate

To the stirred mixture of(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol(600 mg, 2.046 mmol) in DCM (60 mL) was added Et3N (1.426 mL, 10.23mmol), DMAP (50.0 mg, 0.409 mmol) and palmitoyl chloride (1406 mg, 5.11mmol) at 0° C. The reaction mixture was stirred for 3 h at roomtemperature. LCMS showed presence 75% of desired product. The reactionmixture was quenched with water (20 mL) and extracted with DCM (20mL*3). The combined organic layers were washed with water (20 mL), brine(20 mL), dried over Na₂SO₄. After filtration, the mixture wasconcentrated to dryness under vacuum. The residue was purified by silicagel column (120 g, pet. ether:EtOAc=1:1) to afford(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((palmitoyloxy)methyl)tetrahydro-furan-3-ylpalmitate (520 mg) as white solid. The solid was crystallized inpentane:EtOAc=10:1 (10 mL) to give a crystal form, which was dried undersun lamp (45° C.) for 2 h to afford(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((palmitoyloxy)methyl)tetrahydrofuran-3-ylpalmitate (428 mg, 0.546 mmol, 26.7% yield) as while crystal solid. LCMS(ESI) m/z calcd for C₄₄H₇₂FN₅O₅: 770. Found: 793 (M+23)⁺, 771 (M+1)⁺.¹HNMR (300 MHz, CDCl₃) δ 7.97 (s, 1H), 6.42 (t, J=6.3 HZ, 1H), 5.95(brs, 2H), 5.69-5.65 (m, 1H), 4.53-4.39 (m, 2H), 3.04-2.99 (m, 1H),2.75-2.68 (m, 2H), 2.46-2.34 (m, 4H), 1.72-1.61 (m, 4H), 1.42-1.18 (m,48H), 0.99-0.83 (m, 6H).

Example 60:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((icosanoyloxy)methyl)tetrahydrofuran-3-ylicosanoate

To the mixture of icosanoic acid (2.66 g, 8.52 mmol) in DCM (100 mL) wasadded EDC (2.61 g, 13.64 mmol). The reaction mixture was stirred for 30min at 25° C. Then(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydro-furan-3-ol(1.0 g, 3.41 mmol), DMAP (0.833 g, 6.82 mmol)) and DIEA (5.96 mL, 34.1mmol) were added. The reaction mixture was stirred for overnight at 25°C. LCMS showed no desired mass but a new spot was found by TLC (pet.ether:EtOAc=1:1, rf=0.8). The reaction mixture was quenched with water(100 mL), extracted with DCM (50 mL*3). The combined organic layers werewashed with water (100 mL), brine (100 mL), dried over Na₂SO₄. Afterfiltration, the filtrate was concentrated to dryness under vacuum. Theresidue was purified by silica gel (330 g, pet. ether:EtOAc=1:1) toafford(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((icosanoyloxy)methyl)tetra-hydrofuran-3-ylicosanoate (570 mg) as white solid. The solid (500 mg) was dissolved inDCM (250 mL) and MeOH (75 mL), allowed to stand for 3 days at rt andsolids collected by filtration to title compound (260 mg) as irregularcrystals. The solid was dried under sun lamp (45° C.) for 2 h. LCMS:(ESI) m/z calcd for C₅₂H₈₈FN₅O₅: 631. Found: 883 (M+1)⁺, 905 (M+23)⁺.¹HNMR (300 MHz, CDCl₃) δ 7.95 (s, 1H), 6.42 (t, J=6.3 Hz, 1H), 6.05 (brs, 2H), 5.68 (t, J=5.4 Hz, 1H), 4.52 (d, J=12 Hz, 1H), 4.41 (d, J=12 Hz,1H), 3.07-2.98 (m, 1H), 2.78-2.67 (m, 2H), 2.46-2.34 (m, 4H), 1.69-1.60(m, 4H), 1.36-1.23 (m, 64H), 0.90 (t, J=6.3 Hz, 3H).

Example 61:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((undecanoyloxy)methyl)tetrahydrofuran-3-ylundecanoate

To the mixture of(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol(600 mg, 2.046 mmol), Et3N (1.141 mL, 8.18 mmol) and DMAP (50.0 mg,0.409 mmol) in DCM (15 mL) was added undecanoyl chloride (1.3 g, 6.35mmol) dropwise. The reaction mixture was stirred at r.t. for 2 hours.LCMS indicated completion of reaction. The reaction mixture was dilutedwith water (10 mL), extracted with DCM (10 mL×3). The organic phaseswere combined, washed with brine (10 mL), dried over Na₂SO₄, andconcentrated under vacuum. The residue was purified by gel silica column(80 g, EtOAc:pet. ether=1:1) to give the desired product (purity: 97%)as a white solid. The solid was triturated with EtOAc (20 ml×3). Theresulting solid was filtered through a Buchner funnel, rinsed withEtOAc, dried sun lamp (T=50° C.) for 6 h, and was put in a cool dryplace overnight to give(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((undecanoyloxy)methyl)tetrahydro-furan-3-ylundecanoate (560 mg, 99.59%, 42.7% yield) as a white solid. LCMS (ESI)m/z calcd for C₃₄H₅₂FN₅O₅:629. Found: 630 (M+H)⁺. ¹HNMR (400 MHz,Chloroform-d) δ 7.95 (s, 1H), 6.40 (t, J=6.0 Hz, 1H), 6.05 (br, s, 2H),5.65 (dd, J=5.2, 6.8 Hz, 1H), 4.48 (d, J=12.0 Hz, 2H), 4.39 (d, J=12.0Hz, 1H), 3.03-2.96 (m, 1H), 2.74-2.66 (m, 1H), 2.43-2.30 (m, 4H),1.70-1.57 (m, 4H), 1.39-1.18 (m, 28H), 0.90-0.85 (m, 6H).

Example 62:(2R,3aS,26aR)-2-(6-amino-2-fluoro-9H-purin-9-yl)-26a-ethynyldocosahydro-2H-furo[3,2-b][1,5]dioxacyclopentacosine-5,24-dione

To a solution of20-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)-20-oxoicosanoicacid (1.0 g, 1.456 mmol) in ACN (10 mL) and DCM (10 mL) was added TCFH(1.134 g, 4.05 mmol) and NMI (356 mg, 4.86 mmol). The mixture wasstirred at r.t. overnight. The LCMS trace showed the reaction wascompleted. The mixture was concentrated under vacuum and purified bysilica gel (80 g, EtOAc/pet. ether=2/1) to give the crude product (500mg). The crude product was purified by reversed phase (C18, 120 g, A:water (0.05% TFA), B: ACN: 20% to 100% in 60 min, 254/220 nm), thenre-crystallized from (DCM/EtOAc=2/1) to give(2R,3aS,26aR)-2-(6-amino-2-fluoro-9H-purin-9-yl)-26a-ethynyl-docosahydro-2H-furo[3,2-b][1,5]dioxacyclopentacosine-5,24-dione(272.4 mg, 99.5%, yield 27.1%) as a white solid. LCMS (ESI) m/z calcdfor C₃₂H₄₆FN₅O₆: 599 Found: 600 (M+H)⁺. ¹HNMR: (400 MHz, DMSO-d₆) δ8.341 (s, 1H), 7.915-7.894 (brs, 2H), 6.376-6.344 (t, J=6.4 Hz, 1H),5.737-5.709 (t, J=5.6 Hz, 1H), 4.392 (d, J=11.2 Hz, 1H), 4.267 (d,J=11.2 Hz, 1H), 3.713 (s, 1H), 3.172-3.106 (m, 1H), 2.633-2.584 (m, 1H),2.443-2.225 (m, 4H), 1.587-1.533 (m, 4H), 1.266-1.243 (m, 28H).

Example 63:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((hexanoyloxy)methyl)tetrahydrofuran-3-ylhexanoate

To the mixture of(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol(800 mg, 2.73 mmol), Et3N (1.901 mL, 13.64 mmol) and DMAP (167 mg, 1.364mmol) in DCM (20 mL) stirred at room temperature was added hexanoylchloride (1102 mg, 8.18 mmol) dropwise and stirred at for 1 hour. LCMSindicated completion of reaction. The reaction mixture was diluted withwater (10 mL), extracted with DCM (10 mL×3). the organic phases werecombined, washed with brine (10 mL), dried over Na₂SO₄, and concentratedunder vacuum. The residue was purified by gel silica column (80 g,EtOAc: pet. ether=1:1) to give the desired product as a yellow solid.The solid was triturated with EtOAc (10 ml) and stirred for 16 h, theresulting solid was filtered through a Buchner funnel, rinsed withEtOAc, dried beside sun lamp (T=50° C.) for 6 h, and was put in a cooldry place overnight to give(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((hexanoyloxy)methyl)tetra-hydrofuran-3-ylhexanoate (215 mg, 98.05% purity, 15.79% yield) as a white solid. LCMS(ESI) m/z calcd for C₂₄H₃₂FN₅O₅: 489. Found: 490 (M+H)⁺. ¹HNMR (400 MHz,Chloroform-d) δ 8.70 (s, 1H), 6.46 (d, J=5.6 Hz, 1H), 5.51 (d, J=5.2 Hz,1H), 4.55 (d, J=12.0 Hz, 1H), 4.42 (d, J=12.0 Hz, 1H), 2.92 (s, 2H),2.74 (s, 1H), 2.46-2.33 (m, 4H), 1.75-1.55 (m, 6H), 1.40-1.25 (m, 8H),0.97-0.85 (m, 6H).

Example 64:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((nonanoyloxy)methyl)tetrahydrofuran-3-ylnonanoate

To a stirred mixture of(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxy-methyl)tetrahydrofuran-3-ol(500 mg, 1.705 mmol) in DCM (4 mL) was added DMAP (83 mg, 0.682 mmol),triethylamine (863 mg, 8.52 mmol) and nonanoyl chloride (753 mg, 4.26mmol) at 0° C. The reaction mixture was stirred for 6 h at roomtemperature. LCMS showed 49% of desired product. The reaction mixturewas quenched with water (10 mL), extracted with DCM (3*10 mL). Thecombined organic layers were washed with brine and dried over Na₂SO₄.After filtration, the filtrate was concentrated to dryness in vacuum.The residue was purified by silica gel column (40 g, pet.ether:EtOAc=1:1) to afford(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((nonanoyloxy)methyl)tetrahydrofuran-3-ylnonanoate as a yellow oil. Then, the product was re-crystallized fromEtOAc/Hexane in the ratio of 1:1. The solid was collected by filtration,dried under sun lamp (45° C.) to afford(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((nonanoyloxy)methyl)tetrahydrofuran-3-ylnonanoate (262 mg, 0.435 mmol, 25.5% yield) as a white solid. LCMS (ESI)m/z calcd for C₃₀H₄₄FN₅O₅: 573. Found: 574 (M+1)⁺. ¹HNMR (300 MHz,CDCl₃) δ 8.00 (s, 1H), 6.47-6.38 (m, 1H), 6.14-5.80 (m, 2H), 5.64 (t,J=6 Hz, 1H), 4.51-4.38 (m, 2H), 3.04-2.97 (m, 1H), 2.77-2.67 (m, 2H),2.44-2.32 (m, 4H), 1.69-1.50 (m, 4H), 1.48-1.28 (m, 20H), 0.95-0.85 (m,6H).

Example 65:(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((octanoyloxy)methyl)tetrahydrofuran-3-yloctanoate

To a solution of(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxy-methyl)tetrahydrofuran-3-ol (800 mg, 2.73 mmol), Et3N (1.141 mL, 8.18 mmol) andDMAP (66.7 mg, 0.546 mmol) in DCM (8 mL) stirred under nitrogen at 0° C.was added octanoyl chloride (1331 mg, 8.18 mmol) dropwise. The reactionmixture was stirred at 0° C. for 2 h. LCMS indicated completion ofreaction. The reaction mixture was quenched with water and extractedwith DCM (20 mL). The organic phases were washed with water 50 mL, driedover sodium sulphate and evaporated in vacuo to give the crude productsas a yellow oil. The crude product was added to a gel silica column (120g, wet method) and was eluted with Hex/EtOAc (0-30%). Collectedfractions: evaporated in vacuo to give(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((octanoyloxy)methyl)tetra-hydrofuran-3-yloctanoate (233.1 mg, purity: 99.1%, yield: 15.52%) as a white solid. Thesolid in ACN:water=1:1 (5 mL) was stirred at room temperature for 1 hourto obtain crystalline material. LCMS (ESI) m/z calcd for C₂₈H₄₀FN₅O₅:545. Found: 546 (M+1)⁺. ¹HNMR (300 MHz, DMSO-d₆) δ 8.32 (s, 1H), 7.89(s, 2H), 6.34 (t, J=6.6 Hz, 1H), 5.69 (t, J=5.7 Hz, 1H), 4.41 (d, J=11.4Hz, 1H), 4.23 (d, J=11.7 Hz, 1H), 3.78 (s, 1H), 3.32-3.10 (m, 1H),2.65-2.56 (m, 1H), 2.41-2.31 (m, 2H), 1.59-1.45 (m, 4H), 1.28-1.20 (m,16H), 0.92-0.79 (m, 6H).

Example 66:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl3-(2,4-dimethyl-6-(pivaloyloxy)phenyl)-3-methylbutanoate

Step 1: 4,4,5,7-tetramethylchroman-2-one. A mixture of3,5-dimethylphenol (5.0 g, 40.9 mmol) and methyl 3-methylbut-2-enoate(4.67 g, 40.9 mmol) in MeSO₃H (25 mL) was stirred at 70° C. for 6 h.Then, the reaction was cooled, diluted with water (30 mL), extractedwith ethyl acetate (2*20 mL) and the combined organic extracts werewashed with saturated aqueous sodium hydrogencarbonate (2*10 mL) andbrine (50 mL). The mixture was concentrated to dryness under vacuum. Theresidue was purified by silica gel column (80 g, pet. ether:EtOAc=4:1)to afford 4,4,5,7-tetramethylchroman-2-one (8.0 g, 39.2 mmol, 96% yield)as white solid. LCMS (ESI) m/z calcd for C₁₃H₁₆O₂: 204. Found: 246(M+1+ACN)⁺. ¹HNMR (300 MHz, CDCl3) δ: 6.76 (s, 2H), 2.61 (s, 2H), 2.48(s, 3H), 2.29 (s, 3H), 1.46 (s, 6H).

Step 2: 2-(4-hydroxy-2-methylbutan-2-yl)-3,5-dimethylphenol. To asolution of 4,4,5,7-tetramethylchroman-2-one (10 g, 49.0 mmol) in THF(70 mL) was added LiAlH₄ (7.43 g, 196 mmol) in portions at 0° C. Thereaction mixture was stirred for 12 h at room temperature. LCMS showedthe reaction completed. The reaction was quenched with NH₄Cl (sat., aq)at 0° C., extracted with EtOAc (200 mL*3). The combine organic layerswere washed with brine, dried over Na₂SO₄. After filtration, thefiltrate was concentrated to dryness in vacuum and the resulting crudeproduct was used in the next step without purification. LCMS (ESI) m/zcalcd for C₁₃H₂₀O₂: 208. Found: 209 (M+1)⁺.

Step 3:2-(4-((tert-butyldimethylsilyl)oxy)-2-methylbutan-2-yl)-3,5-dimethylphenol.To a solution of 2-(4-hydroxy-2-methylbutan-2-yl)-3,5-dimethylphenol (10g, 48.0 mmol) in DMF (100 mL) was added imidazole (8.17 g, 120 mmol) andTBS-Cl (8.68 g, 57.6 mmol) at 25° C. The reaction mixture was stirredfor 16 h at room temperature. Then, the reaction mixture was quenchedwith water (200 mL), extracted with EtOAc (3×100 mL). The combinedorganic layers were washed with water, brine and dried over Na₂SO₄.After filtration, the filtrate was concentrated in vacuum. The crudeproduct was purified by silica gel (120 g, pet. ether:EtOAc=20:1) toafford2-(4-((tert-butyldimethylsilyl)oxy)-2-methylbutan-2-yl)-3,5-dimethylphenol(9 g, 26.5 mmol, 55.2% yield)) as white solid. LCMS (ESI) m/z calcd forC₁₉H₃₄O₂Si: 322. Found: 323 (M+1)⁺.

Step 4:2-(4-((tert-butyldimethylsilyl)oxy)-2-methylbutan-2-yl)-3,5-dimethylphenylpivalate. To a stirred mixture of2-(4-((tert-butyldimethylsilyl)oxy)-2-methylbutan-2-yl)-3,5-dimethylphenol(3.0 g, 9.30 mmol) in DCM (30 mL) was added Et3N (1.556 mL, 11.16 mmol),DMAP (0.170 g, 1.395 mmol) and pivaloyl chloride (1.346 g, 11.16 mmol)at 0° C. The reaction mixture was stirred for 6 h at room temperature.Then, the reaction mixture was quenched with water (30 mL), organiclayer separated and aqueous layer extracted with DCM (20 mL*3). Thecombined organic layers were washed with water (20 mL), brine (20 mL)and dried over Na₂SO₄. After filtration, the filtrate was concentratedto dryness under vacuum. The residue was purified by silica gel column(80 g, pet. ether:EtOAc=4:1) to afford2-(4-((tert-butyldimethylsilyl)oxy)-2-methylbutan-2-yl)-3,5-dimethylphenylpivalate (3.4 g, 8.36 mmol, 90% yield) as yellow oil. LCMS (ESI) m/zcalcd for C₂₄H₄₂O₃Si: 406. Found: 407 (M+1)⁺.

Step 5: 2-(4-hydroxy-2-methylbutan-2-yl)-3,5-dimethylphenyl pivalate. Toa stirred mixture of2-(4-((tert-butyldimethylsilyl)oxy)-2-methylbutan-2-yl)-3,5-dimethylphenylpivalate (3.4 g, 8.36 mmol) in THF (35 mL) was added water (35.0 mL) andAcOH (100 mL, 1747 mmol) at room temperature. The reaction mixture wasstirred for 6 h at room temperature. Then, the reaction mixture wasquenched with water (50 mL) and extracted with DCM (50 mL*3). Thecombined organic layers were washed with water (20 mL), brine (20 mL)and dried over Na₂SO₄. After filtration, the filtrate was concentratedto dryness under vacuum. The residue was purified by silica gel column(80 g, pet. ether:EtOAc=3:1) to afford2-(4-hydroxy-2-methylbutan-2-yl)-3,5-dimethylphenyl pivalate (2.13 g,7.28 mmol, 87% yield) as yellow oil. LCMS (ESI) m/z calcd for C₁₈H₂₈O₃:292. Found: 293 (M+1)⁺. ¹HNMR (300 MHz, CDCl3). δ 6.82 (s, 1H), 6.43 (s,1H), 3.57 (t, J=6 HZ, 2H), 2.55 (s, 3H), 2.24 (s, 3H), 2.07-2.02 (m,2H), 1.56 (s, 1H), 1.51 (s, 6H), 1.39 (s, 9H).

Step 6:3,5-dimethyl-2-(2-methyl-4-oxobutan-2-yl)phenyl pivalate. To astirred mixture of 2-(4-hydroxy-2-methylbutan-2-yl)-3,5-dimethylphenylpivalate (2.0 g, 6.84 mmol) in DCM (30 mL) was added DMP (5.80 g, 13.68mmol) at 0° C. The reaction mixture was stirred for 3 h at roomtemperature. The reaction mixture was quenched with Na₂S2O₃ (30 ml),NaHCO₃ (30 ml) and extracted with DCM (30 mL*3). The combined organiclayers were washed with Na₂S2O₃ (50 mL), NaHCO₃ (50 mL), brined (50 mL)and dried over Na₂SO₄. After filtration, the filtrate was concentratedto dryness to afford as yellow oil, which was purified by silica gelcolumn (80 g, pet. ether:EtOAc=5:1) to afford3,5-dimethyl-2-(2-methyl-4-oxobutan-2-yl)phenyl pivalate (1.6 g, 5.51mmol, 81% yield) as yellow oil. LCMS (ESI) m/z calcd for C₁₈H₂₆O₃: 290.Found: 308 (M+1+NH₃)⁺.

Step 7: 3-(2,4-dimethyl-6-(pivaloyloxy)phenyl)-3-methylbutanoic acid. Toa stirred mixture of 3,5-dimethyl-2-(2-methyl-4-oxobutan-2-yl)phenylpivalate (1.6 g, 5.51 mmol) in methanol (20 mL) was added water (4.00mL) and oxone (6.77 g, 11.02 mmol) at 0° C. The reaction mixture wasstirred for 6 h at room temperature. The pH was adjusted to 5-6 withsaturated aqueous NaHCO₃. Then reaction mixture was concentrated undervacuum. The water was extracted with DCM (30 mL*3). The combined organiclayers were washed with water (20 mL), brine (20 mL) and dried overNa₂SO₄. After filtration, the filtrate was concentrated to dryness undervacuum. The residue was purified by silica gel column (80 g, pet.ether:EtOAc=4:1) to afford3-(2,4-dimethyl-6-(pivaloyloxy)phenyl)-3-methylbutanoic acid (900 mg,2.94 mmol, 53.3% yield) as white solid. LCMS (ESI) m/z calcd forC₁₈H₂₆O₄: 306. Found: 324 (M+1+NH₃)⁺.

Step 8:((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methyl3-(2,4-dimethyl-6-(pivaloyloxy)phenyl)-3-methylbutanoate. To a solutionof 3-(2,4-dimethyl-6-(pivaloyloxy)phenyl)-3-methylbutanoic acid (400 mg,1.305 mmol), 3-(((ethylimino)methylene)amino)-N,N-dimethyl-propan-1-amine hydrochloride (501 mg, 2.61mmol) in DCM (5 mL) stirred at room temperature was addedN,N-dimethylpyridin-4-amine (31.9 mg, 0.261 mmol) and Et3N (0.546 mL,3.92 mmol) at room temperature. The reaction mixture was stirred at roomtemperature for 1 hr. To the above mixture was added((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenyl-methyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methanol(985 mg, 1.175 mmol). The reaction mixture was stirred at roomtemperature for 16 hr. The reaction mixture was quenched with water (20mL) and extracted with DCM (3*20 mL). The combined organic layers werewashed with brine and dried over Na₂SO₄. After filtration, the filtratewas concentrated to dryness in vacuum. The residue was purified bysilica gel column (80 g, DCM:MeOH=1:1) to afford((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)di-phenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methyl3-(2,4-dimethyl-6-(pivaloyloxy)phenyl)-3-methylbutanoate (1.0 g, 0.888mmol, 68.0% yield) as yellow oil. LCMS (ESI) m/z calcd for C₇₀H₆₈FN₅O₈:1126. Found: 1127 (M+1)⁺.

Step 9:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydro-furan-2-yl)methyl3-(2,4-dimethyl-6-(pivaloyloxy)phenyl)-3-methylbutanoate. To a stirredmixture of((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methyl3-(2,4-dimethyl-6-(pivaloyloxy)phenyl)-3-methylbutanoate (1.0 g, 0.888mmol) in DCM (10 mL) was added TFA (2 mL) at room temperature. Thereaction mixture was stirred for 6 h at room temperature. LCMS showed88% of desired product. The pH of the reaction mixture was adjusted to6-7 with saturated aqueous NaHCO₃ and extracted with DCM (3*10 mL). Thecombined organic layers were washed with brine, dried over Na₂SO₄. Afterfiltration, the filtrate was concentrated to dryness in vacuum. Theresidue was purified by silica gel column (80 g, pet. ether:EtOAc=1:9)to afford((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl3-(2,4-dimethyl-6-(pivaloyloxy)phenyl)-3-methylbutanoate as yellow oil.Then, the product was crystallized from 1:1 EtOAc/Hexane. The solid wascollected by filtration and dried under sun lamp (45° C.) to afford((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl3-(2,4-dimethyl-6-(pivaloyloxy)phenyl)-3-methylbutanoate (224 mg, 0.382mmol, 43.1% yield) as white irregular crystal solid. LCMS (ESI) m/zcalcd for C₃₀H₃₆FN₅O₆: 581. Found: 582 (M+1)⁺. ¹HNMR (300 MHz, CDCl₃) δ7.88 (s, 1H), 6.75 (s, 1H), 6.42 (s, 1H), 6.29-6.25 (m, 1H), 6.02 (br s,2H), 4.27-4.18 (m, 2H), 3.97-3.93 (m, 1H), 3.02 (s, 1H), 2.97 (s, 1H),2.71-2.56 (m, 7H), 2.19 (s, 3H). 1.58 (d, J=6 Hz, 6H), 1.35 (s, 9H).

Example 67:2-(4-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-methyl-4-oxobutan-2-yl)-3,5-dimethylphenyldecanoate

Step 1:2-(4-((tert-butyldimethylsilyl)oxy)-2-methylbutan-2-yl)-3,5-dimethylphenyldecanoate. To a mixture of2-(4-((tert-butyldimethylsilyl)oxy)-2-methylbutan-2-yl)-3,5-dimethylphenol(4.5 g, 13.95 mmol), DMAP (0.170 g, 1.395 mmol) and Et3N (2.333 mL,16.74 mmol) in DCM (45 mL) was added decanoyl chloride (3.19 g, 16.74mmol) at 0° C. The reaction mixture was stirred for 5 hours at roomtemperature. LCMS indicated completion of reaction. The reaction mixturewas quenched with water (50 mL) and extracted with DCM (3*50 mL). Thecombined organic layers were washed with brine and dried over Na₂SO₄.After filtration, the filtrate was concentrated to dryness in vacuum.The residue was purified by silica gel (120 g, pet. ether:EtOAc=8:1) toafford2-(4-((tert-butyldimethylsilyl)oxy)-2-methylbutan-2-yl)-3,5-dimethylphenyldecanoate (5.5 g, 11.53 mmol, 83% yield) as colorless oil. LCMS (ESI)m/z calcd for C₂₉H₅₂O₃Si: 476. Found: 477 (M+H)⁺.

Step 2: 2-(4-hydroxy-2-methylbutan-2-yl)-3,5-dimethylphenyl decanoate.To a mixture of2-(4-((tert-butyldimethylsilyl)oxy)-2-methylbutan-2-yl)-3,5-dimethylphenyldecanoate (3 g, 6.29 mmol) in THF (5 mL) and water (5.00 mL) was addedacetic Acid (15.00 mL) at 25° C. The reaction mixture was stirred for 5hours at 25° C. LCMS indicated completion of reaction. The reactionmixture was quenched with water (30 mL) and extracted with EtOAc (3*30mL). The combined organic layers were washed with NaHCO₃ (aq, 30 mL*2),brine (30 mL) and dried over Na₂SO₄. After filtration, the filtrate wasconcentrated to dryness in vacuum. The residue was purified by gelsilica column (80 g, pet. ether:EtOAc=6:1) to give2-(4-hydroxy-2-methylbutan-2-yl)-3,5-dimethylphenyl decanoate (1.5 g,3.72 mmol, 59.2% yield) as colorless oil. LCMS (ESI) m/z calcd forC₂₃H₃₈O₃: 362. Found: 426 (M+23+ACN)⁺. ¹HNMR (400 MHz, Chloroform-d) δ6.82 (d, J=2.0 Hz, 1H), 6.52 (d, J=2.0 Hz, 1H), 3.54 (t, J=7.2 Hz, 2H),2.57-2.50 (m, 5H), 2.23 (s, 3H), 2.05 (t, J=7.2 Hz, 2H), 1.81-1.69 (m,2H), 1.48 (s, 6H), 1.45-1.23 (m, 12H), 0.92-0.84 (m, 3H).

Step 3: 3,5-dimethyl-2-(2-methyl-4-oxobutan-2-yl)phenyl decanoate. To amixture of 2-(4-hydroxy-2-methylbutan-2-yl)-3,5-dimethylphenyl decanoate(1.5 g, 4.14 mmol) in DCM (30 mL) was added Dess-Martin periodinane(3.51 g, 8.27 mmol) at 25° C. The reaction mixture was stirred for 4hours at 25° C. LCMS indicated completion of reaction. The reactionmixture was quenched with Na₂S2O₃ (aq, 30 mL), NaHCO₃ (aq, 30 mL) andextracted with DCM (3*50 mL). The combined organic layers wereconcentrated to dryness in vacuum. The residue was purified by silicagel (pet. ether:EtOAc=10:1) to afford3,5-dimethyl-2-(2-methyl-4-oxobutan-2-yl)phenyl decanoate (910 mg, 2.272mmol, 54.9% yield) as yellow oil. LCMS (ESI) m/z calcd for C₂₃H₃₆O₃:360. Found: 378 (M+1+NH₃)⁺.

Step 4: 3-(2-(decanoyloxy)-4,6-dimethylphenyl)-3-methylbutanoic acid. Toa mixture of 3,5-dimethyl-2-(2-methyl-4-oxobutan-2-yl)phenyl decanoate(910 mg, 2.52 mmol) in methanol (60 mL) and water (12.00 mL) was addedoxone (3103 mg, 5.05 mmol) at 0° C. The reaction mixture was stirred for12 hours at 25° C. LCMS indicated completion of reaction. The reactionmixture was extracted with ethyl acetate (3*80 mL). The combined organicphases were washed with water and saturated brine, dried over sodiumsulphate and evaporated in vacuum to give the crude product as yellowoil. The residue was purified by gel silica column (80 g, pet.ether:EtOAc=1:1) to give3-(2-(decanoyloxy)-4,6-dimethylphenyl)-3-methylbutanoic acid (450 mg,1.076 mmol, 42.6% yield) as yellow oil. LCMS (ESI) m/z calcd forC₂₃H₃₆O₄: 376. Found: 394 (M+1+NH₃)⁺. ¹HNMR (400 MHz, Chloroform-d) δ6.80 (d, J=2.0 Hz, 1H), 6.56 (d, J=2.0 Hz, 1H), 2.83 (s, 2H), 2.59-2.51(m, 5H), 2.22 (s, 3H), 1.80-1.70 (m, 2H), 1.56 (s, 6H), 1.44-1.22 (m,12H), 0.95-0.85 (m, 3H).

Step 5:2-(4-(((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methoxy)-2-methyl-4-oxobutan-2-yl)-3,5-dimethylphenyldecanoate. To a solution of3-(2-(decanoyloxy)-4,6-dimethylphenyl)-3-methylbutanoic acid (400 mg,1.062 mmol), 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-aminehydrochloride (407 mg, 2.125 mmol) in DCM (5 mL) stirred at room tempwas added N,N-dimethylpyridin-4-amine (260 mg, 2.125 mmol). The reactionmixture was stirred at room temperature for 1 hour. To the above mixturewas added ((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydro-furan-2-yl)methanol(801 mg, 0.956 mmol). The reaction mixture was stirred at roomtemperature for 24 hours. LCMS indicated completion of reaction. Thereaction mixture was quenched with water and extracted withdichloromethane 20 mL. The combined organic phases were washed withsaturated brine 10 mL, dried over sodium sulphate and evaporated invacuum to give the crude product as colorless oil. The residue waspurified by gel silica column (40 g, pet. ether:EtOAc=1:1) to give2-(4-(((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxy-phenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methoxy)-2-methyl-4-oxobutan-2-yl)-3,5-dimethylphenyldecanoate (800 mg, 0.595 mmol, 56.0% yield) as colorless oil. LCMS (ESI)m/z calcd for C₇₅H₇₈FN₅O₈: 1196. Found: 1197 (M+H)⁺. ¹HNMR (400 MHz,Chloroform-d) δ 7.56 (s, 1H), 7.52 (d, J=7.6 Hz, 4H), 7.42-7.34 (m, 2H),7.34-7.15 (m, 17H), 7.00 (s, 1H), 6.84-6.75 (m, 4H), 6.72 (d, J=2.0 Hz,1H), 6.51 (d, J=2.0 Hz, 1H), 6.08 (dd, J=7.6, 4.0 Hz, 1H), 4.51 (d,J=7.6 Hz, 1H), 4.21 (d, J=12.4 Hz, 1H), 3.96 (d, J=12.0 Hz, 1H), 3.77(d, J=18.4 Hz, 6H), 2.78 (s, 1H), 2.71-2.57 (m, 2H), 2.51 (s, 1H), 2.46(td, J=7.6, 3.6 Hz, 2H), 2.39 (s, 3H), 2.28 (s, 1H), 2.19 (s, 3H),1.73-1.63 (m, 2H), 1.43 (s, 3H), 1.36 (s, 3H), 1.28 (s, 6H), 1.24 (s,6H), 0.91-0.83 (m, 3H).

Step 6:2-(4-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-methyl-4-oxobutan-2-yl)-3,5-dimethylphenyldecanoate. To a solution of2-(4-(((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methoxy)-2-methyl-4-oxobutan-2-yl)-3,5-dimethylphenyldecanoate (700 mg, 0.585 mmol) in DCM (10 mL) was added TFA (2 ml, 26.0mmol). The reaction mixture was stirred at room temperature for 1 hr.LCMS indicated completion of reaction. The reaction mixture was quenchedwith water, extracted with dichloromethane 20 mL. The combined organicphases were washed with saturated brine 10 mL, dried over sodiumsulphate and evaporated in vacuum to give the crude product as yellowoil. The residue was purified by reverse phase (C₁₈) column (120 g, FAwith water/MeCN=1:10) to2-(4-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-methyl-4-oxobutan-2-yl)-3,5-dimethylphenyldecanoate (104.3 mg, 0.157 mmol, 26.9% yield) as yellow solid. LCMS(ESI) m/z calcd for C₃₅H₄₆FN₅O₆: 651. Found: 652 (M+H)⁺. ¹HNMR (400 MHz,Chloroform-d) b 8.03 (s, 1H), 6.73 (d, J=2.0 Hz, 1H), 6.54 (d, J=2.0 Hz,1H), 6.27 (dd, J=7.2, 4.4 Hz, 1H), 4.21 (q, J=12.2 Hz, 2H), 3.80 (t,J=7.6 Hz, 1H), 3.11-3.00 (m, 2H), 2.71-2.51 (m, 9H), 2.18 (s, 3H), 1.74(q, J=7.4 Hz, 2H), 1.59 (d, J=18.8 Hz, 6H), 1.43-1.21 (m, 12H),0.92-0.84 (m, 3H).

Example 68:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl(1,3-bis(heptanoyloxy)propan-2-yl) succinate

Step 1: 2-oxopropane-1,3-diyl diheptanoate. To a solution of1,3-dihydroxypropan-2-one (14 g, 155 mmol) in DCM (100 mL) stirred undernitrogen at 0° C. was added pyridine (37.7 mL, 466 mmol), DMAP (0.570 g,4.66 mmol), and heptanoyl chloride (50.8 g, 342 mmol) in DCM (50 mL)dropwise over 15 min. The reaction mixture was stirred at 0° C.overnight. LCMS indicated completion of reaction. The reaction mixturewas quenched with Na₂HCO₃ (100 ml) and the organic layers were washedwith NaCl (100 mL*6), the combined organic layers were dried over Na₂SO₄and evaporated to dryness in vacuo. The residue was purified by flashchromatography (silica gel, 120 g, EtOAc: pet. ether=1:1) to give thedesired product (18 g, yield: 36.8%) as a yellow solid. LCMS (ESI) m/zcalcd for C₁₇H₃₀O₅: 314. Found: 315 (M+1)⁺. ¹HNMR (400 MHz,Chloroform-d): δ 4.77 (s, 4H), 2.46-2.36 (m, 4H), 1.72-1.59 (m, 4H),1.41-1.27 (m, 12H), 0.93-0.89 (m, 6H).

Step 2: 2-hydroxypropane-1,3-diyl diheptanoate. To 2-oxopropane-1,3-diyldiheptanoate (18.0 g, 57.2 mmol) in THF (20 mL) and water (2.000 mL)stirred at 0° C. was added sodium borohydride (3.25 g, 86 mmol)portionwise and stirred at 0° C. for 30 min. LCMS indicated completionof reaction. The reaction mixture was added with water (40 mL),extracted with EtOAc (20 mL*3), the organic phases were combined, washedwith brine (40 mL), dried over Na₂SO₄, and concentrated under vacuum togive crude product (14.8 g, yield: 82%) as white solid which was used inthe next step without purification. LCMS (ESI) m/z calcd for C₁₇H₃₂O₅:316. Found: 317 (M+1)⁺. ¹HNMR (400 MHz, Chloroform-d) b 4.31-4.06 (m,4H), 3.73-3.62 (m, 1H), 2.64-2.43 (m, 1H), 2.39-2.30 (m, 4H), 1.66-1.53(m, 4H), 1.35-1.23 (m, 12H), 0.92-0.86 (m, 6H).

Step 3: 4-((1,3-bis(heptanoyloxy)propan-2-yl)oxy)-4-oxobutanoic acid. Toa solution of 2-hydroxypropane-1,3-diyl diheptanoate (6 g, 18.96 mmol)was dissolved in DCM (30 mL)/THF (30.0 mL) was added pyridine (30.0 mL),DMAP (0.232 g, 1.896 mmol) and dihydrofuran-2,5-dione (3.79 g, 37.9mmol). The resulting mixture was stirred for 6.5 h at 60° C. LCMSindicated completion of reaction. The reaction mixture was diluted withHCl aqueous solution (100 mL), extracted with EtOAc (50 mL*3), theorganic phases were combined, dried over Na₂SO₄, and concentrated undervacuum. The residue was purified by C18 reversed phase column (50-100%ACN/water with 0.1% FA) to give the desired product (7 g, 83.30%, yield:73.8%) as yellow oil. LCMS (ESI) m/z calcd for C₂₁H₃₆O₈: 416. Found: 417(M+1)⁺. ¹HNMR (300 MHz, DMSO-d₆): δ 12.21 (s, 1H), 5.21-5.14 (m, 1H),4.27-4.09 (m, 4H), 2.46-2.41 (m, 4H), 2.32-2.26 (m, 4H), 1.54-1.45 (m,4H), 1.29-1.19 (m, 12H), 0.89-0.82 (m, 6H).

Step 4: 1,3-bis(heptanoyloxy)propan-2-yl(((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methyl)succinate. To a solution of4-((1,3-bis(heptanoyloxy)propan-2-yl)oxy)-4-oxobutanoic acid (596 mg,1.432 mmol) in DMF (6 mL) was added DMAP (525 mg, 4.30 mmol) and3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-aminehydrochloride (824 mg, 4.30 mmol). The resulting mixture was stirred for30 min at room temperature. Then,((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methanol(600 mg, 0.716 mmol) was added and the resulting mixture was stirred forovernight at room temperature. LCMS indicated completion of reaction.The reaction mixture was quenched with water (10 mL), extracted withEtOAc (5 mL*3), the organic phases were combined, washed with brine (10mL), dried over Na₂SO₄, and concentrated under vacuum. The residue waspurified by flash chromatography (silica gel, 80 g, EtOAc:pet.ether=1:1) to give the desired product (470 mg, 97.45%, yield: 51.7%) asa white solid. LCMS (ESI) m/z calcd for C₇₃H₇₈FN₅O₁₂: 1236. Found: 1237(M+1)⁺. ¹HNMR (400 MHz, Chloroform-d) δ 7.67 (s, 1H), 7.54-7.49 (m, 4H),7.41-7.36 (m, 2H), 7.33-7.27 (m, 8H), 7.25-7.16 (m, 10H), 6.81-6.77 (m,4H), 6.10-6.07 (m, 1H), 5.26-5.20 (m, 1H), 4.61-4.54 (m, 1H), 4.34 (d,J=8 Hz, 1H), 4.29-4.24 (m, 2H), 4.16-4.11 (m, 2H), 4.00 (d, J=8 Hz, 1H),3.76 (d, J=16 Hz, 6H), 2.83 (s, 1H), 2.52-2.43 (m, 1H), 2.47-2.34 (m,2H), 2.32-2.26 (m, 4H), 1.63-1.58 (m, 8H), 1.31-1.25 (m, 12H), 0.88-0.83(m, 6H).

Step 5:((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methyl(1,3-bis(heptanoyloxy)propan-2-yl) succinate. To a solution of1,3-bis(heptanoyloxy)propan-2-yl(((2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-(((4-methoxyphenyl)diphenylmethyl)amino)-9H-purin-9-yl)-3-((4-methoxyphenyl)diphenylmethoxy)tetrahydrofuran-2-yl)methyl)succinate (450 mg, 0.364 mmol) in DCM (5 mL) was added TFA (0.561 mL,7.28 mmol) and the resulting mixture was stirred for 1 h at roomtemperature. LCMS indicated completion of reaction. The reaction mixturewas added with MeOH (5 mL), then DCM was removed under vacuum. Themixture was subjected to prep-HPLC (Column: XBridge Prep OBD C18 Column,19*250 mm, 5 um; Mobile Phase A: Water (10 MMOL/L NH4HCO3+0.1% NH3.H2O),Mobile Phase B: ACN; Flow rate: 25 mL/min; Gradient: 65 B to 85 B in 8min; 254 nm; RT1: 6.95;) to give the desired product (29.6 mg, 98.89%,yield: 11.63%) as a white solid. LCMS (ESI) m/z calcd for C₃₃H₄₆FN₅O₁₀:691. Found: 692 (M+1)⁺. ¹HNMR (400 MHz, DMSO) δ 8.27 (s, 1H), 7.84 (br,2H), 6.26-6.23 (m, 1H), 5.78 (d, J=4 Hz, 1H), 5.15-5.13 (m, 1H),5.70-5.64 (m, 1H), 4.39 (d, J=12 Hz, 1H), 4.21-4.09 (m, 5H), 3.62 (s,1H), 2.79-2.73 (m, 1H), 2.56-2.53 (m, 4H), 2.48-2.42 (m, 1H), 2.29-2.25(m, 4H), 1.51-1.35 (m, 4H), 1.37-1.18 (m, 12H), 0.84-0.81 (m, 6H).

Anti-HIV Activity PSV Assay

A pseudotyped virus assay (PSV) was used to assess the potency of theHIV inhibitor. Replication defective virus was produced byco-transfection of a plasmid containing an NL4-3 provirus [containing amutation in the envelope open reading frame (ORF) and a luciferasereporter gene replacing the nef ORF] and a CMV-promoter expressionplasmid containing an ORF for various HIV gp160 envelope clones. Theharvested virus was stored at −80 C in small aliquots and the titer ofthe virus measured to produce a robust signal for antiviral assays.

The PSV assay was performed by using U373 cells stably transformed toexpress human CD4, the primary receptor for HIV entry and either humanCXCR4 or human CCR5 which are the co-receptors required for HIV entry astarget cells for infection. Molecules of interest (including, but notlimited to small molecule inhibitors of HIV, neutralizing antibodies ofHIV, antibody-drug conjugate inhibitors of HIV, peptide inhibitors ofHIV, and various controls) are capable of being diluted into tissueculture media and diluted via serial dilution to create a dose range ofconcentrations, and this was carried out for Example 1. This dose-rangewas applied to U373 cells and the pre-made pseudotyped virus added. Theamount of luciferase signal produced after 3 days of culture was used toreflect the level of pseudotyped virus infection. An IC₅₀, or theconcentration of inhibitor required to reduce PSV infection by 50% fromthe infection containing no inhibitor was calculated. Assays to measurecytotoxity were performed in parallel to ensure the antiviral activityobserved for an inhibitor was distinguishable from reduced target cellviability. IC₅₀ values were determined from a 10 point dose responsecurve using 3-4-fold serial dilution for each compound, which spans aconcentration range >1000 fold. These values are plotted against themolar compound concentrations using the standard four parameter logisticequation:

y=((Vmax*x{circumflex over ( )}n)/(K{circumflex over ( )}n+x{circumflexover ( )}n))+Y2

where:

Y2=minimum y n=slope factor

Vmax=maximum y x=compound concentration [M]

K=EC₅₀

The resulting data is shown in Table 3.

TABLE 3 Example WT IC₅₀ (μM) M184V IC₅₀ (μM) 1 0.002 0.008 2 0.003 0.0133 0.002 0.01 4 0.003 0.017 5 0.001 0.007 6 0.004 0.025 7 0.004 0.024 80.002 0.011 9 0.035 0.167 10 0.009 0.044 11 0.008 0.037 12 0.034 0.15113 0.508 3.61 14 0.033 0.148 15 0.077 0.328 16 0.008 0.031 17 1.74 10.318 0.01 0.073 19 0.018 0.087 20 0.005 0.025 21 0.007 0.044 22 0.0070.035 23 0.011 0.068 24 0.066 0.351 25 0.021 0.186 26 0.007 0.028 270.008 0.046 28 0.006 0.026 29 0.238 1.35 30 0.334 1.5 33 0.017 0.086 340.004 0.025 35 0.001 0.009 36 0.004 0.021

Antiviral Persistence Assay

The PSV assay was adapted to determine the antiviral persistence of eachcompound. This assay evaluates the ability of each compound to remainactive in cells for two days i.e prevent PSV infection of cells in adose dependent manner, 48 h after the removal of compound. Duplicateplates of U373 cells were treated with a serial dilution of smallmolecule inhibitors for 6 h at 37° C. Compounds were removed from cellsby washing twice cells with 1λPBS. For baseline group (i.e immediatelyafter washing or 0 h), cells were infected with prepared PSVs andcultured for three days. For experimental group (48 h), the culturemedium is added to the washed cells and the plate incubated at 37° C.for 48 h. After two days of culture, the prepared PSVs were added to thecells and the mixture cultured for three days. The amount of luciferasesignal produced after culture was used to reflect the level ofpseudotyped virus infection in the baseline group (0 h) and experimentalgroup (48 h) for each compound. An IC₅₀, or the concentration ofinhibitor required to reduce PSV infection by 50% from the infectioncontaining no inhibitor was calculated. The persistence index, which isthe ratio of the IC₅₀ determined at 48 and 0 h is presented in Table 2as well as the fold change of the persistence index relative to EFdA[(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol].

Statistical analysis and graphing of the data were performed in JMP13.2.1 (SAS Institute, Cary, N.C.). A four-parameter logistic Hill Modelwas fit to % Inhibition and log₁₀ concentration values, separately foreach compound, time point and run. A pilot experiment included 2independent experimental runs and a later follow-up experiment included4 runs. Quality control criteria based on R² and 95% confidence intervalranges of all four parameter estimates were used to exclude curves withpoor fits. Using inverse prediction, log₁₀ concentrations were obtainedthat correspond to 50% Inhibition (log₁₀ IC50*) and the log₁₀persistence index was calculated for each compound and run using thefollowing formula: log₁₀ persistence Index=log₁₀ IC50*_(48 hrs)−log₁₀IC50*_(0hrs). Next, a linear mixed effects model was fit on log₁₀persistence index values with a fixed effect for compound and a randomeffect for experimental run, followed by post hoc contrasts to comparethe log₁₀ persistence index of the positive control EFdA to the log₁₀persistence index of other test compounds. The estimated LSMeans anddifferences were then back-transformed via 10^(Estimate) to the originalscale and reported as persistence index and fold change respectively.Raw p-values were reported. Antiviral persistence data for examples15,16,18 and EFdA are shown in Table 4. IC₅₀ curve shifts from t=0 tot=48 h for EFdA and Example 18 are illustrated in FIGS. 1A and 1B. Thecurves in FIG. 1 were obtained from a single curve fit across replicateruns instead of separate fits for each run as described above.

TABLE 4 WT IC₅₀ WT IC₅₀ Fold (μM) at (μM) at Persistence Change Examplet = 0 h t = 48 h Index vs EFdA p-Value EFdA 0.0074 0.3156 42.87 1.00 —15 0.0343 0.1203 3.51 12.23 0.0002 16 0.0055 0.0359 6.49 6.61 0.0042 180.0161 0.0442 2.75 15.59 <0.0001 

Pharmacokinetics Protocol for EFdA Rat PK Studies

A total of seven naïve male Wistar Han rats, 200-250 g, were receivedfrom the supplier equipped with a surgically-implanted jugular veincatheter (JVC) for blood sample collection. Following an acclimationperiod, the animals were assigned to the study based on acceptablehealth as determined by a staff veterinarian and catheter patency.Animals were placed into two groups of 3 rats per group. Fasting of theanimals before or after dosing was not required.

The final study design is presented in the Table 5 below.

TABLE 5 Dose Dose Dose No. of Test Level Conc. Volume Group MalesCompound (mg/kg) (mg/mL) (mL/kg) Dose Vehicle Dose Route 1 3 EFdA 2060.46 0.33 0.5% P407/ Subcutaneous 2 3 20 60.46 0.33 0.5% PEG3350/Intramuscular 3.5% Mannitol in water PEG3350 = Polyethylene glycol 3350.

On Day 1 each animal in Group 1 received a single subcutaneous injectionof prepared test article at a target dose level of 20 mg/kg and at adose volume of 0.33 mL/kg. Each animal in Group 2 received a singleintramuscular injection of prepared test article into a thigh muscle ata target dose level of 20 mg/kg and at a dose volume of 0.33 mL/kg. Theinjection sites were shaved prior to dosing and identified with anindelible marker for daily monitoring during the study. The animals weremanually restrained for dosing and were not sedated. The dose suspensionwas mixed well by inversion before each dose to ensure homogeneity. Alldosing was performed as detailed in the study protocol and was completedwithout incident. Following dosing and at each sample collection timepoint the animals and injection sites were observed for any clinicallyrelevant abnormalities. In addition, all animals are monitored twicedaily by the veterinary staff for clinical abnormalities. No abnormalclinical observations and no injection site reactions were noted duringthe study period.

Blood samples were collected from the study animals as detailed in thesample collection Table 6 below.

TABLE 6 Dose Group/ Collection Information Whole Blood for PK 1 and 20.5, 1, 4, 7, 24, 48, 72 hours post-dose and Day 6, 8, 11, 15, 22, 25,29 Anticoagulant NaF/Na₂EDTA Volume/Time point 250 μL

Each blood sample for PK (250 μL) was collected from the jugular veincatheter or by venipuncture of a jugular vein. The blood was transferredto a tube containing NaF/Na₂EDTA anticoagulant and inverted severaltimes to mix. The blood samples were maintained on crushed ice beforecentrifugation at 2200×g for 10 minutes at 5° C. to isolate plasma. Theresulting plasma samples were transferred to individual polypropylenetubes in a 96-well plate format and immediately placed on dry ice untilstorage at nominally −80° C. prior to analysis.

The PK plasma samples were analyzed by the Testing Facility to determinethe concentration of EFdA using an LC-MS/MS Research Grade Assay (RGA-1)with sensitivity to 0.1 ng/mL. Following review of sample analysis datathrough Day 29, further sample analysis was discontinued and the studywas terminated. The sample concentration data are presented in Tables 7and 8 and mean concentration-time profiles are shown in FIGS. 2 and 3 .

TABLE 7 Plasma concentration of EFdA from rat SC PK study AssayedConcentrations (ng/mL) Time Rat#1 Rat#2 Rat#3 Mean 0.5 hr 1000 640 22101283.33 1 hr 1160 1040 3030 1430.0 4 hr 534 676 708 639.33 7 hr 320 372216 302.67 24 hr 6.70 14.7 2.53 7.98 48 hr 0.771 0.713 0.543 0.68 72 hr0.251 0.270 0.201 0.241 Day 6  BQL BQL 0.117 0.04 Day 8  BQL BQL BQL BQLDay 11 BQL BQL BQL BQL Day 15 BQL BQL BQL BQL Day 22 BQL BQL BQL BQL Day25 0.221 BQL BQL 0.221 Day 29 BQL BQL BQL BQL BQL—Below QuantitationLimit, <0.100 ng/mL

TABLE 8 Plasma concentration of EFdA from rat IM PK study AssayedConcentrations (ng/mL) Time Rat#1 Rat#2 Rat#3 Mean 0.5 hr 929 1150 684921.0 1 hr 957 1640 1400 1332.33 4 hr 662 436 733 610.33 7 hr 241 191426 286.0 24 hr 24.7 1.96 32.6 19.75 48 hr 5.25 0.369 0.915 2.18 72 hr0.301 0.316 0.241 0.29 Day 6  0.102 BQL 0.103 0.10 Day 8  BQL BQL BQLBQL Day 11 BQL BQL BQL BQL Day 15 BQL BQL BQL BQL Day 22 BQL BQL BQL BQLDay 25 BQL BQL BQL BQL Day 29 BQL BQL BQL BQL BQL—Below QuantitationLimit, <0.100 ng/mL

General Protocol for Prodrugs Rat PK Studies

A total of three naïve male Wistar Han rats, 250-300 g, were receivedand following an acclimation period, the animals were assigned to thestudy based on acceptable health as determined by a staff veterinarianand catheter patency. Fasting of the animals before or after dosing wasnot required.

The final study design is presented in the Table 9 below.

TABLE 9 Dose Dose No. of Level Conc. Males (mg/kg) (mg/mL) Dose VehicleDose Route 3 20 2 2% Kolliphor P407 + 2% PEG3350 + Intramuscular(equivalent of EFdA 3.5% Mannitol + 92.5% water PEG3350 = Polyethyleneglycol 3350.

On Day 1, each animal received a single intramuscular injection ofprepared test article at a target dose level of 20 mg/kg equivalent ofEFdA and at a dose volume of 2 mL/kg. The injection sites were atgastrocnemius and identified with an indelible marker for dailymonitoring during the study. The animals were manually restrained fordosing and were not sedated. The dose suspension was mixed well byinversion before each dose to ensure homogeneity. All dosing wasperformed as detailed in the study protocol and was completed withoutincident. Following dosing and at each sample collection time point theanimals and injection sites were observed for any clinically relevantabnormalities. In addition, all animals are monitored twice daily by theveterinary staff for clinical abnormalities. No abnormal clinicalobservations and no injection site reactions were noted during the studyperiod.

Blood samples were collected from the study animals as detailed in thesample collection Table 10 below

TABLE 10 Collection Information Whole Blood for PK Collection time 0.5,1, 3, 5 and 7 hr, 1, 2, 3, 4, 5 and 7 days post dose then 2x/week (Day10 and Day 14, etc) Anticoagulant NaF/Na₂EDTA Volume/Time point 200 μLblood

Each blood sample for PK (200 μL) was collected from tail vein. Theblood was transferred to a tube containing NaF/Na₂EDTA anticoagulant andinverted several times to mix. Draw 150 μL whole blood from the tube,add 150 uL 100 mM ammonium acetate, pH 4 buffer and mix well. Theresulting blood samples were transferred to individual polypropylenetubes and immediately placed on dry ice until storage at nominally −80°C. prior to LC-MS/MS analysis. The mean sample concentration data arepresented in Tables 11 to 31 and mean concentration-time profiles areshown in FIGS. 4 to 24 .

Reference to Example 11

TABLE 11 Plasma concentration of Example 11 and EFdAfrom Example 11 ratIM PK study Dose (mg/kg)/ Sampling Mean concentration Mean concentrationDose route time (day) of Example 11 (ng/mL) of EFdA (ng/mL) 20/IM 0.02086.71 94.0 0.0417 3.37 121 0.125  0.872 55.1 0.208 BQL 38.1 0.292 BQL26.6 1 BQL 5.77 2 BQL 3.90 3 BQL 2.93 4 BQL 3.00 5 BQL 2.85 7 BQL 5.2910 BQL 1.26 14 BQL 1.22 17 BQL 21 BQL 24 BQL 28 BQL 31 BQL 35 BQL 38 BQL42 BQL

TABLE 12 Plasma concentration of Example 17 and EFdA from Example 17 ratIM PK study Dose (mg/kg)/ Sampling Mean concentration Mean concentrationDose route time (day) of Example 17 (ng/mL) of EFdA (ng/mL) 20 0.020815.7 BQL (equivalent 0.0417 28.4 0.900 to EFdA)/IM 0.125 29.1 1.43 0.208 23.0 0.981 0.292 26.7 BQL 1 14.3 BQL 2 2.90 BQL 3 1.47 1.23  41.15 BQL 5 0.99 BQL 7 1.25 BQL 10 1.40 0.878 14 0.757 1.12 

Reference to Example 17

TABLE 13 Plasma concentration of Example 18 and EFdA from Example 18 ratIM PK study Dose (mg/kg)/ Sampling Mean concentration Mean concentrationDose route time (day) of Example 18 (ng/mL) of EFdA (ng/mL) 20 0.020843.1 BQL (equivalent 0.0417 87.0 2.97 to EFdA)/IM 0.125 58.0 4.62 0.20847.1 4.02 0.292 49.2 4.74 1  2.44 BQL 2 BQL 1.91 3 BQL 2.69 4 BQL 2.06 5BQL 2.39 7 BQL 1.88 10 BQL 1.91 14 BQL 2.53 17 BQL 1.90 21 BQL 1.62 24BQL 1.73 28 BQL 1.38 31 BQL 1.56 35 BQL 1.59 38 BQL 1.35 42 BQL BQL 45 0.836 49 BQL 52 BQL 56 BQL 59 BQL 63 BQL 66 BQL

Reference to Example 31

TABLE 14 Plasma concentration of Example 31 and EFdA from Example 31 ratIM PK study Dose (mg/kg)/ Sampling Mean concentration Mean concentrationDose route time (day) of Example 31 (ng/mL) of EFdA (ng/mL) 20 0.02088.33 98.4 (equivalent 0.0417 13.6  370 to EFdA)/IM 0.125 1.65 1107 0.208BQL 669 0.292 BQL 284 1 BQL 46.9 2 6.51 14.7 3 17.3  4.88 4 1.60 2.37 54.05 BQL 7 BQL BQL 10 68.7  BQL 14 BQL BQL 17 BQL BQL 21 BQL BQL 24 BQLBQL 28 BQL BQL 31 BQL BQL

Reference to Example 32

TABLE 15 Plasma concentration of Example 32 and EFdA from Example 32 ratIM PK study Dose (mg/kg)/ Sampling Mean concentration Mean concentrationDose route time (day) of Example 32 (ng/mL) of EFdA (ng/mL) 20 0.020823.2 138 (equivalent 0.0417 20.4 246 to EFdA)/IM 0.125 6.58 185 0.2084.58 102 0.292 2.08 74.2 1 BQL 45.9 2 BQL 41.9 3 BQL 33.8 4 BQL 26.8 5BQL 19.3 7 BQL 3.96 10 BQL 4.28 14 BQL BQL 17 BQL BQL 21 BQL BQL 24 BQLBQL 28 BQL BQL 31 BQL BQL

Reference to Example 37

TABLE 16 Plasma concentration of Example 37 and EFdA from Example 37 ratIM PK study Dose (mg/kg)/ Sampling Mean concentration Mean concentrationDose route time (day) of Example 37 (ng/mL) of EFdA (ng/mL) 20 0.020813.8  31.4 (equivalent 0.0417 13.7  49.3 to EFdA)/IM 0.125 10.0  32.30.208 4.84 25.4 0.292 BQL 17.3 1 BQL 13.7 2 BQL 18.5 3 BQL 13.8 4 BQL11.7 5 7.53 12.2 7 BQL 15.2 10 7.72 12.9 14 BQL 7.13 17 1.82 2.68 211.47 1.76 24 1.25 BQL 28 1.58 BQL 31 6.65 BQL 35 1.70 BQL 38 24.1  BQL42 BQL BQL

Reference to Example

TABLE 17 Plasma concentration of Example 38 and EFdA from Example 38 ratIM PK study Dose (mg/kg)/ Sampling Mean concentration Mean concentrationDose route time (day) of Example 38 (ng/mL) of EFdA (ng/mL) 20 0.020811.2 1.96 (equivalent 0.0417 11.5 3.34 to EFdA)/IM 0.125 14.3 4.30 0.20810.5 4.78 0.292  6.80 3.25 1 BQL 2.31 2 BQL 164 3 BQL 81.7 4  1.83 80.65  2.00 48.9 7 BQL 2.84 10 BQL 3.29 14 BQL 3.75 17 BQL 3.09 21 BQL 3.6324 BQL 3.86 28 BQL 2.55 31 BQL 1.89 35 BQL 2.02 38 1.57 42 1.38 45 1.2549 0.994 52 0.885 56 0.949 59 1.11 63 0.822 66 0.868 70 BQL 73 BQL 77BQL 80 BQL 84 BQL 87 BQL

Reference to Example 39

TABLE 18 Plasma concentration of Example 39 and EFdA from Example 39 ratIM PK study Dose (mg/kg)/ Sampling Mean concentration Mean concentrationDose route time (day) of Example 39 (ng/mL) of EFdA (ng/mL) 20 0.020833.9  69.4 (equivalent 0.0417 36.0  168 to EFdA)/IM 0.125 21.0  1650.208 11.6  156 0.292 7.90 131 1 BQL 121 2 5.52 104 3 BQL 89.2 4 BQL 1055 8.32 109 7 6.13 83.4 10 BQL 39.3 14 BQL 6.92 17 BQL 3.35 21 BQL BQL 24BQL BQL 28 BQL 2.88 31 BQL BQL 35 BQL BQL 38 BQL BQL 42 BQL BQL

Reference to Example 40

TABLE 19 Plasma concentration of Example 40 and EFdA from Example 40 ratIM PK study Dose (mg/kg)/ Sampling Mean concentration Mean concentrationDose route time (day) of Example 40 (ng/mL) of EFdA (ng/mL) 20 0.0208BQL 198 (equivalent 0.0417 BQL 304 to EFdA)/IM 0.125 BQL 288 0.208 BQL210 0.292 BQL 157 1 BQL 51.0 2 BQL 9.36 3 BQL 4.16 4 BQL 1.67 5 BQL BQL7 BQL BQL 10 BQL BQL 14 BQL BQL 17 BQL BQL 21 BQL BQL

Reference to Example 42

TABLE 20 Plasma concentration of Example 42 and EFdA from Example 42 ratIM PK study Dose (mg/kg)/ Sampling Mean concentration Mean concentrationDose route time (day) of Example 42 (ng/mL) of EFdA (ng/mL) 20 0.02082.26 104 (equivalent 0.0417 BQL 148 to EFdA)/IM 0.125 BQL 117 0.208 BQL94.2 0.292 BQL 93.2 1 BQL 35.6 2 BQL 13.3 3 BQL 6.00 4 BQL BQL 5 BQL BQL7 BQL BQL 10 BQL BQL 14 BQL BQL 17 BQL BQL 21 BQL BQL 24 BQL BQL 28 BQLBQL 31 BQL BQL

Reference to Example 45

TABLE 21 Plasma concentration of Example 45 and EFdA from Example 45 ratIM PK study Dose (mg/kg)/ Sampling Mean concentration Mean concentrationDose route time (day) of Example 45 (ng/mL) of EFdA (ng/mL) 20 0.020835.6 30.1 (equivalent 0.0417 49.1 90.3 to EFdA)/IM 0.125 42.8 181 0.20815.7 98.5 0.292 6.15 61.4 1 0.847 23.1 2 BQL 16.9 3 BQL 12.7 4 BQL 6.535 1.56 6.96 7 BQL 5.59 10 BQL 5.64 14 BQL BQL 17 BQL BQL 21 BQL BQL 24BQL 28 BQL 31 BQL 35 BQL

Reference to Example 48

TABLE 22 Plasma concentration of Example 48 and EFdA from Example 48 ratIM PK study Dose (mg/kg)/ Sampling Mean concentration Mean concentrationDose route time (day) of Example 48 (ng/mL) of EFdA (ng/mL) 20 0.020810.4 BQL (equivalent 0.0417 15.9 1.43 to EFdA)/IM 0.125 7.04 2.32 0.2084.43 2.13 0.292 2.00 2.51 1 BQL 4.11 2 BQL 6.57 3 BQL 10.3 4 BQL 14.3 5BQL 17.7 7 BQL 10.8 10 BQL 1.86 14 BQL BQL 17 BQL BQL 21 BQL BQL

Reference to Example 50

TABLE 23 Plasma concentration of Example 50 and EFdA from Example 50 ratIM PK study Dose (mg/kg)/ Sampling Mean concentration Mean concentrationDose route time (day) of Example 50 (ng/mL) of EFdA (ng/mL) 20 0.0208BQL 3223 (equivalent 0.0417 BQL 3077 to EFdA)/IM 0.125 BQL 239 0.208 BQL38.7 0.292 BQL 19.3 1 BQL 1.48 2 BQL BQL 3 BQL BQL 4 BQL BQL 5 BQL BQL 7BQL BQL 10 BQL BQL 14 BQL BQL 17 BQL BQL 21 BQL BQL

Reference to Example 58

TABLE 24 Plasma concentration of Example 58 and EFdA from Example 58 ratIM PK study Dose (mg/kg)/ Sampling Mean concentration Mean concentrationDose route time (day) of Example 58 (ng/mL) of EFdA (ng/mL) 20 0.020811.4 1.20 (equivalent 0.0417 14.9 1.10 to EFdA)/IM 0.125 9.18 1.02 0.2087.95 BQL 0.292 7.82 BQL 1 2.96 BQL 2 1.05 BQL 3 0.844 BQL 4 BQL BQL 5BQL  0.838 7 BQL 1.34 10 BQL 1.62 14 1.94 17 2.10 21 BQL 24 1.22 28 1.2331  0.879 35 BQL 38 BQL 42 BQL 45 BQL 49 BQL

Reference to Example 59

TABLE 25 Plasma concentration of Example 59 and EFdA from Example 59 ratIM PK study Dose (mg/kg)/ Sampling Mean concentration Mean concentrationDose route time (day) of Example 59 (ng/mL) of EFdA (ng/mL) 20 0.020820.8 BQL (equivalent 0.0417 26.7  0.899 to EFdA)/IM 0.125 43.3 2.290.208 42.4 2.08 0.292 42.4 BQL 1 8.72 BQL 2 4.23 BQL 3 2.52 BQL 4 2.36BQL 5 BQL 7 BQL 10 2.66 14 2.31

Reference to Example 61

TABLE 26 Plasma concentration of Example 61 and EFdA from Example 61 ratIM PK study Dose (mg/kg)/ Sampling Mean concentration Mean concentrationDose route time (day) of Example 61 (ng/mL) of EFdA (ng/mL) 20 0.020893.3 BQL (equivalent 0.0417 189 2.21 to EFdA)/IM 0.125 64.3 2.63 0.20888.7 2.02 0.292 87.4 1.63 1 13.6 BQL 2 BQL BQL 3 BQL BQL 4 BQL BQL 5 BQLBQL 7 BQL BQL 10 BQL BQL 14 BQL BQL 17 BQL BQL 21 BQL BQL

Reference to Example 62

TABLE 27 Plasma concentration of Example 62 and EFdA from Example 62 ratIM PK study Dose (mg/kg)/ Sampling Mean concentration Mean concentrationDose route time (day) of Example 62 (ng/mL) of EFdA (ng/mL) 20 0.020826.7 5.50 (equivalent 0.0417 27.6 17.2 to EFdA)/IM 0.125 8.98 34.8 0.2084.98 27.7 0.292 4.79 24.5 1 1.47 8.31 2 0.839 6.42 3 0.887 5.51 4 1.575.93 5 2.08 5.43 7 2.36 5.06 10 1.82 3.94 14 1.68 2.23 17 BQL BQL 21 BQLBQL 24 BQL BQL 28 BQL BQL

Reference to Example 63

TABLE 28 Plasma concentration of Example 63 and EFdA from Example 63 ratIM PK study Dose (mg/kg)/ Sampling Mean concentration Mean concentrationDose route time (day) of Example 63 (ng/mL) of EFdA (ng/mL) 20 0.02083.28 183 (equivalent 0.0417 1.92 386 to EFdA)/IM 0.125 BQL 176 0.208 BQL98.0 0.292 BQL 75.1 1 BQL 43.6 2 BQL 23.0 3 BQL 12.7 4 BQL 5.64 5 BQL3.63 7 BQL 2.23 10 BQL BQL 14 BQL BQL 17 BQL 21 BQL 24 BQL 28 BQL

Reference to Example 64

TABLE 29 Plasma concentration of Example 64 and EFdA from Example 64 ratIM PK study Dose (mg/kg)/ Sampling Mean concentration Mean concentrationDose route time (day) of Example 64 (ng/mL) of EFdA (ng/mL) 20 0.02081.30 5.48 (equivalent 0.0417 2.05 19.9 to EFdA)/IM 0.125  0.994 22.10.208 BQL 16.6 0.292 BQL 9.58 1 BQL 8.61 2 BQL 7.63 3 BQL 8.31 4 1.078.99 5 BQL 13.2 7 BQL 13.3 10 BQL 4.70 14 BQL 2.98 17 2.38 21 1.42 24BQL 28 BQL

Reference to Example 65

TABLE 30 Plasma concentration of Example 65 and EFdA from Example 65 ratIM PK study Dose (mg/kg)/ Sampling Mean concentration Mean concentrationDose route time (day) of Example 65 (ng/mL) of EFdA (ng/mL) 20 0.02082.75 24.7 (equivalent 0.0417 4.33 105 to EFdA)/IM 0.125 9.69 130 0.208BQL 88.3 0.292 BQL 61.0 1 BQL 35.1 2 BQL 29.5 3 BQL 17.1 4 BQL 15.0 5BQL 10.5 7 BQL 2.51 10 BQL 14 BQL 17 BQL 21 BQL 24 BQL 28 BQL

Reference to Example 66

TABLE 31 Plasma concentration of Example 66 and EFdA from Example 66 ratIM PK study Dose (mg/kg)/ Sampling Mean concentration Mean concentrationDose route time (day) of Example 66 (ng/mL) of EFdA (ng/mL) 20 0.02081.51 322 (equivalent 0.0417 0.928 480 to EFdA)/IM 0.125 BQL 209 0.208BQL 135 0.292 BQL 117 1 BQL 44.7 2 BQL 22.5 3 BQL 13.3 4 BQL 4.83 5 BQL3.78 7 BQL 1.25 10 BQL 14 BQL 17 BQL 21 BQL 24 BQL 28 BQL

Protocol for Example 18 Rat PK Studies

Example 18 referenced herein was made in accordance with Synthesis B. Atotal of seven naïve male Wistar Han rats, 250-275 g, were received fromthe supplier equipped with a surgically-implanted jugular vein catheter(JVC) to facilitate blood sample collection during the first severaldays of the study. Following an acclimation period, the animals wereassigned to the study based on acceptable health as determined by astaff veterinarian and catheter patency. Six animals were placed intotwo groups of 3 rats per group. Fasting of the animals before or afterdosing was not required.

The final study design is presented in the Table 32 below.

TABLE 32 Dose Dose Dose No. of Level Conc. Volume Group Males TestArticle (mg/kg) (mg/mL) (mL/kg) Dose Vehicle Dose Route 1 3 Example 1820 84.53 0.49 1% P407/ Subcutaneous (prodrug) (prodrug) 1% PEG 335041.19 in PBS (parent, EFdA) 2 3 20 84.53 0.49 Intramuscular (prodrug)41.19 (parent, EFdA) PEG = polyethylene glycol; PBS = phosphate-bufferedsaline

On Day 1 each animal in Group 1 received a single intrascapularsubcutaneous injection of prepared test article at a target dose levelof 20 mg parent/kg and at a dose volume of 0.49 mL/kg. Each animal inGroup 2 received a single intramuscular injection of prepared testarticle into a thigh muscle at a target dose level of 20 mg parent/kgand at a dose volume of 0.49 mL/kg. The animals were manually restrainedfor dosing and were not sedated. The dose sites were clipped of hair andwiped with alcohol before dosing. All dosing was performed as detailedin the study protocol and was completed without incident.

Following dosing and at least twice daily until the end of the study,the animals and dose sites were observed for any clinically relevantabnormalities. All animals appeared normal at the time of eachobservation and no dose site reactions were observed Blood samples werecollected from the study animals as detailed in the sample collectionTable 33 below.

TABLE 33 Group/ Whole Blood for PK Collection Information 1A and 2A 0.5,1, 3, 5, 7, 24, 48, 72 hours post-dose (N = 3 per group) and Day 8, 11,15, 22, 25, 29, 32, 36, 39, 43, 46, 50, 53, 57, 60, 64, 67, 71, 74, 78,81, 85, 88, 92 Anticoagulant NaF tubes containing Na₂EDTA Volume/Timepoint 250 μL of whole blood

Interim blood samples for PK (Groups 1A and 2A) were collected from thejugular vein catheter or by venipuncture of a jugular vein if thecatheter was not patent. Blood was transferred to a blood collectiontube containing NaF/Na₂EDTA anticoagulant, mixed by inversion, andmaintained on wet ice until processing. The blood samples werecentrifuged at 2200×g for 10 minutes at 5° C. to isolate plasma. Theresulting plasma samples were transferred to individual polypropylenetubes in a 96-well plate format and immediately placed on dry ice untilstorage at nominally −80° C. prior to analysis. The PK plasma sampleswere analyzed every 1-2 weeks by the Testing Facility to determine theconcentration of prodrug and parent using an LC-MS/MS Research GradeAssay (RGA-1) with sensitivity to 0.1 ng/mL. Following interim review ofsample analysis data, analysis for prodrug was discontinued after theDay 29 sample collections. The sample concentration data are presentedin Tables 34 and 35, and mean concentration-time profiles are shown inFIGS. 25 and 26

TABLE 34 Plasma concentration of Example 18 and EFdA from Example 18 ratSC PK study SC Dosing 20 mg/kg equivalent of EFdA Time, Assayed Example18 Concentrations Assayed EFdA Concentrations Day Rat#1 Rat#2 Rat#23Mean Rat#1 Rat#2 Rat#3 Mean 0.021 7.40  BQL BQL 7.40 0.464 0.194 0.2200.29 0.042 3.88  0.103 BQL 1.99 0.819 0.337 0.349 0.50 0.125 4.19  0.1570.143 1.50 1.63 0.644 0.701 0.99 0.208 1.80  0.155 0.200 0.72 1.74 0.8700.768 1.13 0.292 0.804 0.127 0.150 0.36 1.61 0.770 0.704 1.03 1 0.211BQL BQL 0.21 1.38 0.792 0.855 1.01 2 BQL BQL BQL BQL 1.11 0.631 0.5150.75 3 BQL BQL BQL BQL 1.18 0.570 0.499 0.75 8 0.158 BQL  0.156 0.161.12 0.979 0.874 0.99 11 BQL BQL BQL BQL 1.32 1.28 0.949 1.18 15 0.101BQL BQL 0.10 1.50 1.84 1.41 1.58 22 BQL BQL BQL BQL 0.787 0.971 1.070.94 25 BQL BQL BQL BQL 1.03 1.10 0.913 1.01 29 BQL BQL BQL BQL 0.6811.27 0.833 0.93 32 0.533 0.905 0.742 0.73 36 0.573 0.668 0.726 0.66 390.504 0.811 0.630 0.65 43 0.546 0.728 0.768 0.68 46 0.399 0.603 0.6520.55 50 0.715 0.850 1.13 0.90 53 0.696 0.614 1.04 0.78 57 0.475 0.6310.770 0.63 61 0.520 0.589 0.517 0.54 64 0.359 0.567 0.742 0.56 67 0.3180.476 0.454 0.42 71 0.315 0.524 0.546 0.46 74 0.673 0.476 0.473 0.54 780.379 0.373 0.490 0.41 81 0.392 0.491 0.896 0.59 85 0.719 0.423 1.800.98 88 0.338 0.407 0.465 0.40 92 0.666 0.484 0.689 0.61

TABLE 35 Plasma concentration of Example 18 and EFdA from Example 18 ratIM PK study IM Dosing 20 mg/kg equivalent of EFdA Time, Assayed Example18 Concentrations Assayed EFdA Concentrations Day Rat#1 Rat#2 Rat#23Mean Rat#1 Rat#2 Rat#3 Mean 0.021 0.111 15.9 0.233 5.41 0.279 0.4050.299 0.33 0.042 0.620 11.5 0.408 4.18 0.734 0.779 1.01 0.84 0.125 0.6891.45 0.314 0.82 1.37 1.67 1.62 1.55 0.208 0.594 3.26 0.340 1.40 1.552.05 2.09 1.90 0.292 0.377 8.80 0.239 3.14 1.41 1.75 1.46 1.54 1 0.3250.276 BQL 0.30 1.28 1.22 1.41 1.30 2 BQL BQL BQL BQL 1.15 1.21 1.35 1.243 BQL BQL BQL BQL 1.33 1.12 1.12 1.19 8 0.133 0.156 BQL 0.14 1.45 1.221.30 1.32 11 0.130 0.102 BQL 0.12 1.58 0.864 1.48 1.31 15 0.130 BQL BQL0.13 1.35 1.24 1.28 1.29 22 BQL BQL BQL BQL 1.59 0.974 1.11 1.22 25 BQLBQL BQL BQL 1.23 0.934 1.45 1.20 29 BQL BQL BQL BQL 0.976 1.01 1.27 1.0932 0.789 0.608 0.920 0.77 36 0.688 0.588 0.857 0.71 39 0.869 0.690 0.8630.81 43 0.869 0.919 0.865 0.88 46 0.875 0.595 0.825 0.77 50 1.10 0.8681.25 1.07 53 0.690 0.756 0.724 0.72 61 0.571 0.903 0.923 0.80 64 0.5240.457 0.785 0.59 67 0.648 0.457 1.04 0.72 71 0.468 0.597 1.03 0.70 740.435 0.400 0.704 0.51 78 0.486 0.410 0.841 0.58 81 0.563 0.444 0.6220.54 85 0.434 0.704 0.622 0.59 88 0.419 0.465 0.586 0.49 92 0.734 0.6281.02 0.79

Protocol for Example 18 (Compound) Dog PK Studies

Example 18 referenced herein was made in accordance with Synthesis B.Six male beagle dogs (+1 spare) were selected from the TestingFacility's colony of non-naïve animals. The animals were assigned to thestudy based on acceptable health as determined by a Testing Facilityveterinarian. Six animals were selected and placed into two groups ofthree animals each. Fasting of the animals before or after dosing wasnot required.

On Day 1 each animal in Group 1 received a single intrascapularsubcutaneous injection of prepared test article at a target dose levelof 5 mg parent/kg in 2% P404/2% PEG3350/PBS (dose concentration 97.36mg/mL prodrug) and at a dose volume of 0.1 mL/kg. Each animal in Group 2received a single intramuscular injection of prepared test article in ahind quarter (quadriceps) at a target dose level of 5 mg parent/kg in 2%P404/2% PEG3350/PBS (dose concentration 97.36 mg/mL prodrug) and at adose volume of 0.1 mL/kg. The animals were manually restrained fordosing and were not sedated. The dose sites were clipped of hair andwiped with alcohol before dosing. All dosing was performed as detailedin the study protocol and was completed without incident. Followingdosing and at least twice daily until the end of the study, the animalsand dose sites were observed for any clinically relevant abnormalities.All animals appeared normal at the time of each observation and no dosesite reactions were observed. Blood samples were collected from thestudy animals as detailed in the sample collection Table 26 below.

TABLE 36 Group/ Whole Blood for PK Collection Information 1-2 0.5, 1, 3,5, 7, 24, 48, 72 hours post-dose and Day 8, 10, 17, 19, 22, 25, 29, 32,36, 39, 43, 46, 50, 53, 57, 60, 64, 67, 71, 74, 78, 81, 85, 88, 92Anticoagulant NaF tubes containing Na2EDTA Volume/Time point 500 μL ofwhole blood

Blood samples were collected by venipuncture of a cephalic vein. For PK,blood was transferred to a blood collection tube containing NaF/Na₂EDTAanticoagulant, mixed by inversion, and maintained on wet ice untilprocessing. The blood samples were centrifuged at 2200×g for 10 minutesat 5° C. to isolate plasma. The resulting plasma samples weretransferred to individual polypropylene tubes in a 96-well plate formatand immediately placed on dry ice until storage at nominally −20° C.prior to shipment for analysis.

The PK plasma samples were analyzed approximately every two weeks by theTesting Facility to determine the concentration of Example 18 (prodrug)and parent using an LC-MS/MS Research Grade Assay (RGA-1) withsensitivity to 0.1 ng/mL. Following interim review of sample analysisdata, analysis for Example 18 (prodrug) was discontinued after the Day57 sample collections. The sample concentration data are presented inTables 37 and 38, and mean concentration-time profiles are shown inFIGS. 27 and 28

TABLE 37 Plasma concentration of Example 18 and EFdA from Example 18 DogSC PK study SC Dosing 20 mg/kg equivalent of EFdA Time Example 18Assayed Concentrations (ng/mL) EFdA Assayed Concentrations (ng/mL)(days) Dog1001 Dog1002 Dog1003 Mean Dog1001 Dog1002 Dog1003 Mean 0.021BQL BQL BQL BQL BQL BQL BQL BQL 0.042 BQL 0.176 BQL 0.176 BQL BQL BQLBQL 0125 0.205 2.00 0.461 0.889 0.110 0.638 0.174 0.307 0.208 0.673 1.941.33  1.31  0.204 1.11 0.628 0.647 0.292 1.11  2.11 1.74  1.65  0.4461.44 1.22 1.04 1 4.19  3.62 5.59  4.47  1.80 2.17 2.90 2.29 2 1.14  1.181.06  1.13  1.54 1.38 1.86 1.59 3 0.906 0.931 0.747 0.861 1.70 1.56 2.041.77 8 0.281 0.215 0.600 0.365 1.93 1.72 2.11 1.92 10 0.111 0.171 0.2640.182 1.79 1.46 2.04 1.76 17 BQL 0.106 BQL 0.106 0.916 0.942 1.37 1.0819 BQL BQL BQL BQL 0.824 0.904 0.935 0.888 22 0.116 BQL BQL 0.116 0.8690.699 1.40 0.989 25 BQL BQL BQL BQL 0.588 0.434 0.702 0.575 29 BQL BQL0.101 0.101 0.555 0.556 0.747 0.619 32 BQL BQL 0.108 0.108 0.622 0.4710.791 0.628 36 BQL BQL BQL BQL 0.533 0.440 0.738 0.570 39 BQL BQL BQLBQL 0.550 0.660 0.353 0.521 43 BQL BQL BQL BQL 0.433 0.372 0.582 0.46246 BQL BQL BQL BQL 0.364 0.319 0.526 0.403 50 BQL BQL BQL BQL 0.2510.251 0.295 0.266 53 BQL BQL BQL BQL 0.363 0.345 0.457 0.388 57 BQL BQLBQL BQL 0.265 0.244 0.249 0.253 60 0.265 0.246 0.283 0.265 64 0.1870.222 0.280 0.230 67 0.189 0.229 0.260 0.226 71 0.203 0.185 0.240 0.20974 0.259 0.277 0.422 0.319 78 0.229 0.235 0.350 0.271 81 0.238 0.1640.324 0.242 85 0.194 0.214 0.250 0.219 88 0.146 0.123 0.190 0.153 920.152 0.170 0.124 0.149 BQL—Below Quantitation Limit, <0.100 ng/mL

TABLE 38 Plasma concentration of Example 18 and EFdA from Example 18 dogIM PK study IM Dosing 20 mg/kg equivalent of EFdA Time Example 18 plasmaconcentrations (ng/mL) EFdA plasma concentrations (ng/mL) (days) Dog2001Dog2002 Dog2003 Mean Dog2001 Dog2002 Dog2003 Mean 0.021 2.43 2.67 1.472.19 0.158 0.344 BQL 0.251 0.042 3.38 2.85 1.90 2.71 0.682 1.51 0.5660.919 0125 10.5 7.99 4.36 7.62 3.47 3.92 2.30 3.23 0.208 9.34 7.61 7.518.15 3.79 4.17 3.61 3.86 0.292 8.41 7.29 4.96 6.89 3.07 3.81 3.41 3.43 12.45 2.69 2.03 2.39 2.56 3.67 2.57 2.93 2 1.04 1.21 1.19 1.15 1.92 2.982.02 2.31 3 1.09 0.877 0.818 0.928 1.92 2.18 2.23 2.11 8 0.204 0.4190.201 0.275 2.10 2.16 2.20 2.15 10 0.129 0.198 0.150 0.159 1.46 1.882.05 1.80 17 0.110 0.172 0.116 0.133 1.25 1.51 1.17 1.31 19 0.109 BQLBQL 0.109 0.864 1.14 0.961 0.988 22 BQL 0.113 BQL BQL 0.997 1.25 1.271.17 25 BQL BQL BQL BQL 0.492 0.675 0.681 0.616 29 BQL BQL BQL BQL 0.7340.679 0.927 0.780 32 BQL BQL BQL BQL 0.581 0.739 0.688 0.669 36 BQL BQLBQL BQL 0.643 0.575 0.702 0.640 39 BQL BQL BQL BQL 0.677 0.600 0.5900.622 43 BQL BQL BQL BQL 0.441 0.465 0.506 0.471 46 BQL BQL BQL BQL0.519 0.464 0.527 0.503 50 BQL BQL BQL BQL 0.229 0.244 0.284 0.252 53BQL BQL BQL BQL 0.387 0.444 0.395 0.409 57 BQL BQL BQL BQL 0.218 0.2440.244 0.235 60 0.290 0.262 0.360 0.304 64 0.272 0.215 0.219 0.235 670.308 0.244 0.277 0.276 71 0.171 0.246 0.217 0.211 74 0.372 0.273 0.4190.355 78 0.364 0.285 0.269 0.306 81 0.239 0.214 0.335 0.263 85 0.2390.201 0.302 0.247 88 0.182 0.165 0.210 0.186 92 0.111 0.137 0.132 0.127BQL—Below Quantitation Limit, <0.100 ng/mL

Protocol for Example 18 (Compound) Cynomolgus Monkey PK Study

Example 18 referenced herein was made in accordance with Synthesis B.Six male cynomolgus monkeys (+1 spare) were selected from the TestingFacility's colony of non-naïve animals. The animals were assigned to thestudy based on acceptable health as determined by a Testing Facilityveterinarian. Six animals were selected and placed into two groups ofthree animals each. Fasting of the animals before or after dosing wasnot required.

On Day 1 each animal in Group 1 received a single intrascapularsubcutaneous injection of prepared test article at a target dose levelof 5 mg parent/kg in 2% P404/2% PEG3350/PBS (dose concentration 97.36mg/mL prodrug) and at a dose volume of 0.1 mL/kg. Each animal in Group 2received a single intramuscular injection of prepared test article in ahind quarter (quadriceps) at a target dose level of 5 mg parent/kg in 2%P404/2% PEG3350/PBS (dose concentration 97.36 mg/mL prodrug) and at adose volume of 0.1 mL/kg. The animals were manually restrained fordosing and were not sedated. The dose sites were clipped of hair andwiped with alcohol before dosing. All dosing was performed as detailedin the study protocol and was completed without incident. Followingdosing and at least twice daily until the end of the study, the animalsand dose sites were observed for any clinically relevant abnormalities.All animals appeared normal at the time of each observation and no dosesite reactions were observed. Blood samples were collected from thestudy animals as detailed in the sample collection Table 39 below.

TABLE 39 Group/ Whole Blood for PK Collection Information 1-2 0.5, 1, 3,5, 7, 24, 48, 72 hours post-dose and Day 8, 10, 17, 19, 22, 25, 29, 32,36, 39, 43, 46, 50, 53, 57, 60, 64, 67, 71, 74, 78, 81, 85, 88, 92,,106, Anticoagulant NaF tubes containing Na2EDTA Volume/Time point 500 μLof whole blood

Blood samples were collected by venipuncture of a cephalic vein. For PK,blood was transferred to a blood collection tube containing NaF/Na₂EDTAanticoagulant, mixed by inversion, and maintained on wet ice untilprocessing. The blood samples were centrifuged at 2200×g for 10 minutesat 5° C. to isolate plasma. The resulting plasma samples weretransferred to individual polypropylene tubes in a 96-well plate formatand immediately placed on dry ice until storage at nominally −20° C.prior to shipment for analysis.

The PK plasma samples were analyzed approximately every two weeks by theTesting Facility to determine the concentration of Example 18 (prodrug)and parent using an LC-MS/MS Research Grade Assay (RGA-1) withsensitivity to 0.1 ng/mL. Following interim review of sample analysisdata, analysis for Example 18 (prodrug) was discontinued after the Day57 sample collections. The bioanalytical report and sample concentrationdata are presented in Tables 40 and 41, and mean concentration-timeprofiles are shown in FIGS. 29 and 30

TABLE 40 Plasma concentration of Example 18 and EFdA from Example 18monkey SC PK study SC Dosing 20 mg/kg equivalent of EFdA Example 18plasma concentrations (ng/mL) EFdA plasma concentrations (ng/mL) TimeMonkey Monkey Monkey Monkey Monkey Monkey (days) 1001 1002 1002 Mean1001 1002 1003 Mean 0.021 49.4 0.337 0.133 0.235 BQL BQL BQL BQL 0.04270.3 1.63 0.179 0.905 0.126 BQL BQL 0.126 0.125 149 5.59 2.29 3.9400.492 0.241 0.561 0.431 0.208 92.8 7.99 1.91 4.950 0.767 0.721 0.8940.794 0.292 146 6.67 2.38 4.525 1.13 0.738 0.970 0.946 1 36.5 1.60 0.6411.121 2.27 1.06 1.49 1.607 2 0.734 0.230 0.579 0.405 1.56 1.12 1.851.510 3 0.694 0.228 0.396 0.312 1.62 1.27 1.56 1.483 8 BQL 0.102 0.1310.117 1.79 1.37 1.45 1.537 10 0.146 0.111 0.146 129 1.64 1.28 1.57 1.49717 BQL BQL BQL BQL 1.84 1.35 1.51 1.567 19 BQL BQL BQL 2.57 1.40 1.701.890 22 BQL BQL BQL 2.71 1.59 1.68 1.993 25 BQL BQL BQL 2.16 1.47 1.921.850 29 BQL BQL BQL 1.66 1.20 1.66 1.507 32 BQL BQL BQL 1.94 1.23 1.671.613 36 BQL BQL BQL 1.82 1.05 1.50 1.457 39 BQL BQL BQL 1.56 1.00 1.491.350 43 BQL BQL BQL 1.36 0.929 1.52 1.270 46 BQL BQL BQL 1.21 0.9351.22 1.122 50 BQL BQL BQL 1.36 0.802 1.08 1.081 53 BQL BQL BQL 0.8950.741 0.908 0.848 57 BQL BQL BQL 1.18 0.639 0.693 0.837 60 1.12 0.7030.739 0.854 64 1.02 0.721 0.847 0.863 67 0.845 0.670 0.921 0.812 71 1.020.569 0.682 0.757 74 1.32 0.876 1.08 1.092 78 1.16 0.899 1.01 1.023 811.39 0.795 0.744 0.976 85 1.16 0.490 0.744 0.798 88 0.957 0.527 0.4840.656 92 0.803 0.608 0.527 0.646 106 0.713 0.692 0.696 0.700 120 0.5640.430 0.538 0.511 BQL—Below Quantitation Limit, <0.100 ng/mL

TABLE 41 Plasma concentration of Example 18 and EFdA from Example 18monkey IM PK study IM Dosing 20 mg/kg equivalent of EFdA Example 18plasma concentrations (ng/mL) EFdA plasma concentrations (ng/mL) TimeMonkey Monkey Monkey Monkey Monkey Monkey (days) 2001 2002 2003 Mean2001 2002 2003 Mean 0.021 21.8 40.2 1.93 11.865 0.135 BQL BQL 0.1350.042 29.2 82.6 0.790 14.995 0.843 0.317 0.258 0.473 0.125 53.7 363 3.3128.505 1.59 1.15 0.745 1.162 0.208 51.3 692 5.27 28.285 3.57 1.66 1.262.163 0.292 30.1 821 6.42 18.260 2.44 2.44 1.79 2.223 1 6.89 110 2.084.485 4.24 4.15 5.01 4.467 2 0.652 0.674 0.387 0.520 3.35 2.93 2.422.900 3 0.379 0.533 0.630 0.505 2.86 2.77 2.24 2.623 8 0.203 0.248 0.1710.187 4.03 3.34 2.31 3.227 10 0.156 0.251 0.190 0.173 2.99 3.02 2.502.837 17 0.135 0.160 0.154 0.145 4.10 2.28 2.90 3.093 19 0.119 0.1490.129 0.124 2.72 2.04 2.30 2.353 22 0.146 0.114 0.113 0.130 2.21 1.992.53 2.243 25 BQL 0.125 0.151 0.151 2.18 1.85 3.70 2.577 29 0.155 0.135BQL 4.39 2.90 1.87 3.053 32 BQL BQL BQL 3.13 1.61 1.80 2.180 36 BQL BQLBQL 1.56 1.55 1.65 1.587 39 BQL BQL BQL 1.92 1.51 1.41 1.613 43 BQL BQLBQL 1.27 1.29 1.57 1.377 46 BQL BQL 0.105 1.29 1.19 1.36 1.280 50 BQLBQL BQL 1.16 1.16 1.24 1.187 53 BQL BQL BQL 1.42 1.77 1.40 1.530 57 BQLBQL BQL 1.20 0.791 1.05 1.014 60 1.91 1.95 1.14 1.667 64 1.24 0.9740.984 1.066 67 1.23 1.36 0.817 1.136 71 1.84 0.712 0.861 1.138 74 1.321.15 1.28 1.250 78 1.12 1.09 1.10 1.103 81 1.21 1.18 1.02 1.137 85 0.8351.01 0.890 0.912 88 1.02 0.750 0.626 0.799 92 0.699 0.582 0.724 0.668106 0.855 0.828 0.816 0.833 120 0.643 0.595 0.589 0.609 BQL—BelowQuantitation Limit, <0.100 ng/mL

As shown above pharmacokinetic results, in rat IM and SC administrationof 20 mg/kg EFdA showed a detectable duration of exposure of EFdA forless than 8-days, likely reflecting rapid absorption of the compound asa consequence of its high solubility and high permeability. Therefore,an approach involving design of novel lipophilic prodrugs of EFdA as ameans to modulate the physicochemical properties of the EFdA as astrategy to improve its suitability for use as a long-acting injectableantiviral agent was pursued. The rat, dog and monkey IM and SC PK datademonstrate that sustained exposure of EFdA for extended period wasachieved with several lipophilic prodrugs suggests potential utility oflipophilic prodrugs in HIV therapy as a long-acting IM or SCadministrative prodrugs of EFdA.

The specific pharmacological responses observed may vary according toand depending on the particular active compound selected or whetherthere are present pharmaceutical carriers, as well as the type offormulation and mode of administration employed, and such expectedvariations or differences in the results are contemplated in accordancewith practice of the present disclosure.

1-32. (canceled)
 33. A compound having the structure:

or a pharmaceutically acceptable salt thereof.
 34. A pharmaceuticalcomposition comprising a compound according to claim 32, or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable excipient.
 35. The composition of claim 34, wherein thecomposition is present in parenteral form.
 36. The composition of claim34, wherein the composition is in a tablet form.
 37. The composition ofclaim 34, wherein the composition is formulated as a long actingparenteral injection.
 38. A method of treating an HIV infection in apatient comprising administering to the subject a compound of claim 33,or a pharmaceutically acceptable salt thereof.
 39. A compound having thestructure:


40. A pharmaceutical composition comprising a compound according toclaim 39, and a pharmaceutically acceptable excipient.
 41. Thecomposition of claim 40, wherein the composition is present inparenteral form.
 42. The composition of claim 40, wherein thecomposition is in a tablet form.
 43. The composition of claim 40,wherein the composition is formulated as a long acting parenteralinjection.
 44. A method of treating an HIV infection in a patientcomprising administering to the subject a compound of claim 39.